KR20150000878A

KR20150000878A - Methods and apparatuses harvesting, modifying and reimplantation of dermal micro-organs

Info

Publication number: KR20150000878A
Application number: KR1020147024971A
Authority: KR
Inventors: 스테펜 에프. 벨로모; 알렉스 오쿤; 야론 퓨르스트; 다비드 샬헤베트; 엘리샤 아미르; 모르데차이 버크만
Original assignee: 메드제닉스 메디칼 이스라엘 리미티드
Priority date: 2012-02-09
Filing date: 2012-11-28
Publication date: 2015-01-05
Also published as: JP2015507962A; EP2811924A1; CA2864237A1; CN104114111A; AU2012369064A2; AU2012369064A1; HK1204899A1; AU2012369064A8; MX2014009507A

Abstract

Embodiments of the present invention provide methods and apparatus for harvesting dermal micro-organ (DMO). Some embodiments of the present invention provide a DMO comprising a plurality of dermal components that substantially maintains the microstructure and three-dimensional structure of the dermis tissue from which it originated. According to some embodiments of the present invention, the DMO sampling device 5000, 6000 includes a support structure for supporting the skin-related tissue structure from which the DMO is to be harvested, a cutting tool 3014 for separating the DMO from the skin- , 6016). Exemplary embodiments of the invention provide genetically modified dermal micro-organisms that express one or more recombinant gene products. Some embodiments of the present invention provide methods and apparatus for transgenic DMO transplantation.

Description

Field of the Invention [0001] The present invention relates to a method and apparatus for harvesting, deforming and re-implanting dermal micro-

The present invention relates to a method and apparatus field for collecting, processing, transplanting and manipulating tissue-based micro-organs, tissue-based therapeutic micro-organs and dermal tissues.

Various methods for delivering therapeutic agents are known. For example, the therapeutic agent can be delivered orally, transdermally, by inhalation, by infusion, and by sustained release. In each of these cases, the delivery method is limited by the in-vivo process through which the therapeutic agent is administered, the need for frequent administration, and the size limitations of the molecules that can be used. In some methods, the amount of the therapeutic agent varies from administration to administration.

Dermal micro-organ (DMO) can be maintained autonomously for a long period of time outside the body ("in vitro" or "in vitro"), and various operations can be applied, It may be implanted subcutaneously or for the purpose of treating a disorder or for plastic surgery purposes. The DMO may be modified to express the gene product of interest. This modified dermal micro-organ is generally referred to as a therapeutic dermal micro-organ (DTMO).

Skin micro-organs, including epidermal and dermal tissue layers, have been observed in a variety of clinical trials, as outlined schematically in PCT / IL02 / 0880, for example. When taking a skin sample, there will be a superficial wound that lasts for several weeks to leave a scar. The sampled skin samples require significant processing to produce micro-organs from these samples. In addition, keratinocysts or keratinocysts are known to occur when subcutaneous or subcutaneous deep-tissue transplantation of the skin micro-organ. In addition, when implanting skin micro-organs such as implants in a "slit" manner on the skin surface, a significant technical expert is needed to handle the MO while maintaining proper orientation.

For example, techniques for harvesting dermis to be used as "filler material " in cosmetic surgery or cosmetic procedures are known in the art. Conventional harvesting techniques include epidermal stripping techniques using a scalpel or surgical scalpel to expose a dermal section. Such skin draining or surgical scalpels can be used again to manually take the exposed dermis fragments.

Other conventional devices for collecting dermis include, but are not limited to, Martin Dermal (Part No. P-225) available from Padgett (Part No. P-225) for use in harvesting dermal cores from the back, There is a Harvester. In order to operate this device which is not commonly used, a sharp cutting tube containing a reusable tube having an inner diameter of about 4.5 mm and a thick wall must be manually rotated at a very slow speed. Generally, in order to use this type of device, pressure is applied to the skin surface just above the harvesting site, and the suture is squeezed while pushing the cutting tube forward. In addition, the dermis thus collected is generally not uniform in size and includes a skin "plug" at both ends of the dermal core.

Embodiments of some aspects of the present invention provide a DMO / DTMO having the ability to be maintained in a viable state normally in vitro, as disclosed in U.S. Patent Application Publication No. US-2012/0201793-A1 , Enabling various manipulations on DMO, while maintaining a high level of production and secretion of the desired therapeutic agent, the entire contents of which are incorporated herein by reference. Embodiments of some aspects of the present invention also include methods for obtaining a DMO and then transplanting the DTMO without implanting DTMO into the keratin sac or keratin microcaps when implanted subcutaneously or deep into the body, for example, The method comprising the steps of: In addition, since the method and apparatus according to some embodiments of the present invention are relatively simple, the level of skill required by an expert in performing the method of the present invention and / or using the apparatus of the present invention, It will be understood by one of ordinary skill in the art that the level required to do so is not as high.

Some embodiments of the present invention provide a dermis micro-organ (DMO) having a plurality of dermis components that may comprise cells of the dermis tissue and surrounding substrates. The DMO according to embodiments of the present invention typically maintains the micro-architecture and tertiary structure of the dermis organs originating from DMO, and the dimensions of the DMO are due to malnutrition and accumulation of waste products May be of a size that permits passive diffusion of the appropriate nutrients and gases into the cell and diffusion of cellular waste outside the cell to minimize cytotoxicity and thus death.

In some embodiments of the present invention, the DMO of the present invention does not produce keratin, or produces keratin at a negligible level.

In some embodiments of the invention, the DMO does not produce keratin and / or keratin cysts after subcutaneous implantation or implantation into the deep region of the body.

In another embodiment of the present invention, the DMO of the present invention produces a keratinocyst that is atrophied within a relatively short period of time, e. G., Days or weeks after subcutaneous implantation.

In another embodiment of the invention, the DMO comprises a hair follicle and sebaceous glands.

Further, exemplary embodiments of the present invention provide methods and apparatus for harvesting dermal micro-organ. The method may include, for example, stabilizing and / or supporting a skin-related tissue structure from which the dermis micro-organ is harvested, such that the skin-related tissue structure can be maintained in a desired shape and / or location; Separating at least DMO from the skin-associated tissue structure; And separating the DMO extracted from the body. According to some of these embodiments, the support structure includes a vacuum chamber, which can maintain the skin-associated tissue structure in a desired shape and / or position, such that the DMO can be cut from the skin- . In one embodiment, the support structure includes at least one vacuum channel for fluidly coupling a vacuum chamber with one or more vacuum sources.

In one embodiment, the harvesting apparatus for dermal micro-organ comprises: (a) a support structure for supporting a tissue structure associated with the skin from which the DMO is to be harvested; (b) an introducer; And (c) a cutting tool, the support structure comprising a first tubular element, the first tubular member including an insert into the apparatus. In some embodiments, the first tubular member is a guide channel that can guide additional members, e.g., a cutting tool, for insertion into a skin-related tissue that is supported.

In one embodiment, the apparatus of the present invention further comprises a vacuum chamber comprising: (a) two proximal protrusions, one with proximal protrusions and one with distal protrusions, adjacent to the insert, elevated protrusion; And (b) a central channel located between the two raised protrusions, wherein the raised protrusions comprise at least a plateau comprising a skin layer and a dermis layer derived from a skin-associated tissue structure, Wherein the central channel is configured to support a portion of the skin epidermal layer derived from the skin-associated tissue structure and the layer of skin-derived tissue derived from the skin-related tissue structure, such that the dermal layer of the skin is located in the orbit of the cutting tool when the cutting tool is inserted into the first tubular member of the device. Supports the dermal layer.

In another embodiment, the apparatus of the present invention comprises a introducer comprising a second tubular member and a fourth tubular member, the second tubular member being inserted into the fourth tubular member and passing through the proximal end of the fourth tubular member And the second tubular member and the fourth tubular member are coaxially inserted into the first tubular member at the insertion portion together; Further, upon withdrawal of the second tubular member, the fourth tubular member is coaxially left in the first tubular member.

In another embodiment, the apparatus includes a third tubular member, e.g., a cutting tool, inserted into and passing through the fourth tubular member.

In one embodiment, the cutting tool comprises a coring tube that is capable of cutting through a skin-related tissue structure when advanced along a cutting axis, the cutting axis being substantially coaxial with the first tubular member . In another embodiment, the core ring tube is a rotatable core ring tube attached to a power source.

In one embodiment, the vacuum chamber comprises a vacuum control mechanism. An implementation of a vacuum state may include, for example, placing a finger in a vacuum chamber hole, for example, a vacuum hole, which, when covered, forms a vacuum. As another example, releasing the vacuum state may include releasing the finger from the vacuum hole. As another example, covering or uncovering a vacuum hole can be used to control and release the vacuum state, respectively. In one embodiment, the vacuum control mechanism is governed by opening or closing the valve of the vacuum control line, or by clamping or unclamping the vacuum line.

In one embodiment, the DMO harvesting method of the present invention comprises the steps of placing a device in the harvesting section in contact with the surface of the individual's skin; Supporting the tissue-related tissue structure to be harvested in the DMO; Puncturing the skin related tissue structure; Cutting the DMO from the supported skin-related tissue structure; And recovering the DMO. In another embodiment, the harvesting method comprises making only one perforation in the skin-related structure.

In one embodiment, the method of collection includes recovering DMO from a coring tube to a closed vessel using a vacuum. In one embodiment, the enclosed container is a syringe body. In one embodiment, the syringe may have an attached bulkhead. In another embodiment, when the coring tube is withdrawn from the collection section, the DMO is present in the coring tube and the recovery of the DMO comprises flushing the DMO from the coring tube.

Further, exemplary embodiments of the present invention provide methods and apparatus for transplanting dermal micro-organ. In one embodiment, the dermal micro-organ to be implanted is a genetically modified dermis micro-organ, which may also be referred to herein as a therapeutic dermal micro-organ (DTMO).

In one embodiment, a DMO or DTMO implantation device comprises: (a) a loading syringe comprising a first tubular member; (b) an implantation tool comprising a second tubular member; (c) a support structure for securing the skin-associated tissue structure to a location where the DMO is to be implanted into the skin-associated tissue structure; (d) a introducer for perforating the skin in the infiltration part; (e) a stopper tool connectable to the support structure, the stopper tool including a tubular member, the stopper tool assisting in maintaining the position of the DMO during withdrawal of the implant tool .

In an exemplary embodiment of the invention, the DMO implantation method comprises the steps of: (a) loading a DTMO into a loading syringe comprising a first tubular member; (b) moving the DTMO from the loading syringe to an implantable tool including a second tubular member; (c) disposing an implantation device in the implant, the device being in contact with a skin layer of the subject, the implantation axis being generally parallel to the skin layer; (d) supporting the skin-associated tissue structure at the implant to implant the DTMO into a skin-associated structure; (e) perforating the skin within the skin-associated tissue structure at the infiltration site, the method comprising: drilling using a introducer comprising an inner needle and an outer sleeve member; (f) removing the inner needle of the introducer and advancing the implant tool along the implantation axis to the skin related tissue structure; And (g) withdrawing the second tubular member, leaving the DTMO in the skin-associated tissue structure. In one embodiment, a stopper tool is used to assist in maintaining the position of the DTMO during withdrawal of the implant. In one embodiment, the first and second steps may be optional since the DTMO can be directly loaded into the distal portion of the implant needle by sucking the DTMO from the back end of the needle using a syringe.

In other embodiments, DTMO may be implanted by direct injection of the DTMO from the syringe through the needle under the skin, under the skin or at other anatomical locations, if linear implantation is not important.

Further, exemplary embodiments of the present invention are directed to genetically modified dermal micro-organisms that express one or more recombinant gene products, that is, dermal tissue cells that maintain the micro-structure and three-dimensional structure of the dermal tissue from which it originated And matrices and allows passive diffusion of nutrients and gases into cells and diffusion of extracellular cell waste and minimization of cytotoxicity and subsequent death due to malnutrition and accumulation of waste products Wherein at least a portion of the cells of the DMO comprise at least one recombinant gene product or at least one recombinant gene product as disclosed in U.S. Publication No. US-2012-0201793-A1. And more particularly, the entirety of which is incorporated herein by reference. In another exemplary embodiment, the at least one recombinant gene product is at least one recombinant protein.

In some embodiments of the invention, the genetically modified DMO of the present invention does not substantially produce keratin.

In some embodiments, the invention provides a method of delivering recombinant gene products produced by DMO to a recipient.

In some embodiments, the present invention provides a method of inducing a physiological local or systemic effect by implanting DMO into a recipient.

In another embodiment, the invention provides a method of delivering a protein of interest to a subject. The method includes transplanting genetically modified DMO into the skin, under the skin, or at other locations in the body.

In another embodiment, the present invention provides a DTMO transplantation method for avoiding or alleviating keratinocaps formation.

In one embodiment, the present invention provides a method of removing implanted DTMO.

BRIEF DESCRIPTION OF THE DRAWINGS [0029] For a reading of the present disclosure, non-limiting embodiments of the invention are described in the following description. In the drawings, identical or similar structures, elements or parts thereof, which appear in more than one illustration, are generally designated by the same or similar reference in the drawings in which they are represented. The sizes of the components and features shown in the drawings are mainly selected for convenience and clarity, and accumulation is not necessarily accurate.
1 is a schematic block diagram illustrating an example of a method for producing and using a therapeutic dermal micro-organ (DTMO) according to an exemplary embodiment of the present invention.
2 is a schematic flow chart describing a DMO collection method in accordance with some exemplary embodiments of the present invention.
Figures 3A-3G schematically illustrate exemplary DMO harvesting steps in accordance with the method of Figure 2.
Figures 4A-4E illustrate implementations for a harvesting device, a medical drill for use with a harvesting device, a syringe for use in harvesting DTMO, bulkheads, and some members of a collect. 4A shows an example of the harvest syringe 4002 and the partition 4004. 4B is an example of a collector. 4C is an example of a support structure in which a vacuum hole is covered with a finger. 4D shows one embodiment of the introducer: an inner needle (4006-needle) and an outer guide (4008-white sleeve). 4E illustrate one embodiment of a cutting tube 4010, a drill 4012, and a drill handpiece 4014. [
5A-B schematically illustrate some of the components of a dermal sampling device according to another exemplary embodiment of the present invention. Figure 5A schematically illustrates a side view of harvesting device implementations. Figure 5B schematically shows a cross-sectional view of the device of Figure 5A supporting the skin-related tissue structure from outside which is capable of harvesting dermal micro-organ at the desired location.
Figure 6 schematically illustrates some of the components of dermal sampling devices according to another exemplary embodiment of the present invention.
7 is a flow chart describing a method of implanting a DTMO in accordance with some exemplary implementations of the present invention.
Figures 8A-E illustrate one embodiment of an implant device. 8A is an example of an embodiment of a loading syringe, 8B is an embodiment of an implantation tool, 8C is an embodiment of a introducer, 8D is an embodiment of a support structure, and 8E is an embodiment of a stopper.
9A-E schematically illustrate exemplary steps of transplanting a DTMO according to the method of FIG.
10A-B illustrate implementations of a syringe having a bulkhead and a collector. 10A shows a syringe having a bulkhead and a collector inserted through a guide channel and an outer sleeve of a support structure, wherein the support structure is connected to a vacuum source. Fig. 10B shows an embodiment of a syringe with a collect and a needleless valve attached to the back of the coring needle. The T-shaped end of the introducer 1008 is indicated for directional use.
Figure 11 schematically illustrates one embodiment of a support structure.
FIG. 12 is a flowchart describing a method of removing the already-implanted DTMO.
13 shows one embodiment of a syringe 1306 attached to a glued-on male luercap 1302 near the posterior portion of the coring needle 1304. As shown in Fig.
14A and 14B show examples of a dermal micro-organ (Fig. 14A) and a collection unit 1402 and a transplant unit 1404 of a human subject.
It will be appreciated that for simplicity and clarity of illustration, elements shown in the figures are not necessarily drawn to scale. For example, the size of some of the elements may be exaggerated relative to other elements for clarity. In addition, where considered appropriate, reference to a number may be repeated in the figures to indicate the same or similar elements.

In the following detailed description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. It will be apparent, however, to one skilled in the art that the present invention may be practiced without these specific details. In other instances, well known methods, processes, and components have not been described in detail so as not to obscure the present invention.

It is to be understood that the following description is given in terms of specific uses and needs, and that those skilled in the art will be able to make and use the present invention. Various modifications to the described implementations will be apparent to those skilled in the art, and the principles defined herein may be applied to other implementations. Accordingly, the invention is not to be limited to the specific embodiments shown and described herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein. In yet another case, the methods, processes and components described in U.S. Patent Publication No. US-2012/0201793-A1 are incorporated herein by reference in their entirety.

I. An exemplary definition of the terms used herein

As used herein, the term "explant " refers, in some embodiments of the invention, to a truncated fragment of a living tissue or organ derived from one or more tissues or organs of an individual, For example, it may be a dermal micro-organ.

As used herein, the term "dermis micro-organ" or "DMO" is used in some embodiments of the present invention to assist in the viability and function of cells, while at least partially preserving similar in vivo interactions with the tissues or organs obtained Quot; means a separate tissue or organ structure derived from a meal or produced in a manner that results in a meal or meal. DMOs can contain multiple dermal components that maintain the microstructure of the tissue or organ from which they are derived and the tertiary structure of the dermis tissue from which they originate and are also useful for the prevention and treatment of cytotoxicity due to malnutrition and waste accumulation, , A size selected to allow passive diffusion of appropriate nutrients and gases into DMO inner cells. The DMO may consist essentially of a plurality of dermal components (skin tissue components located under the epidermis). These components may include dermal fibroblasts, epithelial cells, other cell types, follicular bases, nerve endings, sebaceous glands, glands, blood vessels, and lymphatic vessels. In the following, the description of the embodiment for DMO also relates to MO. Further, when the term "dermal tissue" is used, it also applies to the "dermal tissue"

In some embodiments herein, the term "microstructure" is used herein to refer to at least about 50%, in other embodiments at least about 60%, in yet another embodiment at least about 70% , In yet another embodiment about 80% or more, and in yet another embodiment about 90% or more, of a cell or non-cellular material having physical and / or functional contact in vivo, &Lt; / RTI > and / or maintaining a functional contact. Preferably, the exhaled bacterium cells possess one or more biological activities of the organ or tissue from which they are isolated.

The term "donor " is used herein to refer to an entity, in some embodiments of the present invention, that takes an eating segment and uses it to form one or more micro-organs, or one or more micro-organ forms already exist. In one embodiment, the donor is a human individual. In other embodiments, the donor is a mammalian entity other than human.

As used herein, the term " therapeutic micro-organ (TMO) "is used in some embodiments of the present invention to treat a disease or condition, such as a mutated or modified DMO to produce a therapeutic agent, Means a dermal micro-organ (MO) that can be used to promote the purpose. The therapeutic substance may or may not be a natural biomaterial. In all of the following cases, the discussion of DTOM implementations also applies to DTMO, which is a therapeutic dermis MO that can be genetically modified in some embodiments of the invention.

As used herein, the term "transplantation " means, in some embodiments of the invention, introducing one or more TMOs or DTMOs into a recipient, wherein the TMO or DTMO is derived from a recipient organism, It may be from an animal tissue. TMO or DTMO can be transplanted slit into the skin, transplanted subcutaneously, or implanted into a desired site in the body of the recipient body by substitution. In one embodiment, the DTMO is derived from the recipient tissue. In one embodiment, the DTMO is substantially implanted into the dermal layer of the dermal tissue. In one embodiment, the DTMO is implanted between the dermis and the fat layer of the skin tissue.

As used herein, the term " male body " means, in some embodiments of the invention, an individual to which one or more TMOs or DTMOs are implanted. In one embodiment, the recipient is a human. In another embodiment, the recipient is a mammalian entity other than human. In some embodiments, the receptor is provided by one or more autologous TMOs or DTMOs.

As used herein, the term "in vitro" should be understood to include "ex-vivo ".

As used herein, the term "coring tube ", individually or collectively, refers to any element having similar functionality as well as the terms" cutting tool ", " Lt; / RTI > In some embodiments, the coring needle of the present invention is disposable.

As used herein, the term "implantation tool " may relate, individually or collectively, to the terms" implant needle "," implant needle "and" implant tube "as well as any other element having a similar function. In some embodiments, the implantable device of the present invention is disposable.

As used herein, the term "tubular member " refers to a member having or consisting of a tube, wherein the tube generally refers to any structure or device of various cylinders. In one embodiment, the tubular member is a member having a shape of a hollow elongated cylinder. In another embodiment, the tubular member is a member having a tunnel or channel shape that cuts through a cylindrical channel, such as a solid mass. In another embodiment, the tubular member is open at both ends. In another embodiment, the tubular member is open at one end. In another embodiment, the tubular member includes a beveled needle tip at one end. In other embodiments, the tubular member includes at least one sharp blunt end. In one embodiment, the tubular member is a solid, non-hollow member having a long cylindrical shape, for example a rod. In one embodiment, the tubular member may comprise a guide channel.

As used herein, the term "rod" refers to a linear three-dimensional member having a solid geometry. In one embodiment, the rod has a circular cross-section. In one embodiment, the rod has a non-circular cross-section.

As used herein, the term "skin related tissue structure" refers to a tissue, tissue, or tissue that can be stabilized and / or supported by an apparatus as defined herein to allow dermal micro- Refers to the structure of the components. Skin-related tissue structures may include epidermal tissue components and dermal tissue components. Alternatively, the skin-associated tissue structure may include adipose tissue and / or muscle tissue present in the vicinity of the dermal tissue.

In one embodiment, the skin-associated tissue construct of the present invention comprises skin tissue components that have been drawn into the central channel under vacuum conditions. In one embodiment, the skin-associated structure comprises epidermis, dermis and fatty tissue. In other embodiments, the skin-related constructs include epidermis, dermis, fats, and muscle tissue.

Herein, the term "center channel" is used interchangeably with the term "vacuum chamber" in some embodiments of the present invention.

As used herein, the term "coaxial" refers to radial symmetry consisting of components positioned about the same axis or about the same axis. In this way, the tubular members can be positioned approximately equidistant from the common axis. In one embodiment, the cutting tool is arranged approximately equidistantly relative to the common axis presented by the guide channel. In one implementation, the cutting tools are arranged approximately equidistantly relative to the common axis presented by the center channel.

As used herein, the term "approximately" refers to a range of values that belong to + or -10% in an ideal result. For example, substantially coaxial tubular members may have the same central axis, or may have central axes located within 10% of the same common central axis.

In one embodiment, one tubular member is contained within another tubular member, but the central axes of these tubular members need not be in line.

On the other hand, for clarity and completeness of explanation, all aspects of the production and use of DTMO are described herein, and while embodiments of the invention describe from start to end of the process, May be used with other methods and / or devices for performing other aspects, and it is to be understood that portions described herein may be used for other purposes. The present invention includes portions for the production and maintenance of dermal micro-organ for transformation into DTMO. It should be understood that the dermal micro-organ produced according to these aspects of the present invention may also be used for purposes other than transformation into DTMO.

In some embodiments of the invention, the micro-organ comprises a plurality of dermal components, such as fibroblasts, and / or nerve endings, sebaceous glands, glands, blood vessels, lymphatics, elastin fibers and / One epidermal constituent, and / or an endothelial component, and / or an immune system derived cell, and / or an extracellular matrix. When a micro-organ containing an existing epidermal layer ("split thickness skin MO") is routinely subcutaneously implanted with a mouse and a pig, a keratin sac or giant-keratin sac can be formed. In contrast, according to exemplary embodiments of the present invention, when the DMO is sampled using the skin tissue as a sample, or DMO is directly sampled, it can be used as a mouse, a pig, or a human, even after transplanted or anatomically transplanted No cysts or giant sacs are found in humans. The biological activity of DTMO according to embodiments of the present invention (e. G., Secretion of therapeutic proteins such as erythropoietin and resulting rise in hematocrit) may be similar to or higher than that in TMO derived from partial skin .

Generally, the production of DTMOs can include DMO harvesting steps, DMO maintenance steps and / or DMO modification steps and / or genetically modified steps thereof, and in some embodiments, the desired material (e.g., protein) Lt; RTI ID = 0.0 > production. &Lt; / RTI > The use of DTMO may include the production of a therapeutic substance, e. G. Protein, in the body of a patient or an animal, in order to treat the individual. For example, DTMOs can be transplanted into the skin, under the skin, or into the body of an individual to produce a substance / protein in vivo.

In one embodiment, the DTMO is not encapsulated in an immunocompromising capsule or shell.

In some embodiments of the invention, the DMO may contain tissue of the basal epidermal layer and optionally other skin layers of the skin. In another embodiment, the dermal micro-organ does not include underlying epidermal tissue.

In some embodiments of the present invention, the DMO does not include skin layers. In other embodiments, the DMO may comprise several layers of epidermal tissue. In some embodiments, the dermal micro-organ may lack a complete skin layer. In certain cases, the DMO may involve the incorporation of epidermis in dermal tissue layers, but still the complete epidermal layer may be absent.

In one embodiment of the invention, the DMO comprises an entire cross-section of the dermis. In another embodiment of the invention, the dermal micro-organ comprises a portion of the dermal cross-section. In another embodiment, the DMO comprises most of the transverse segments of the dermis, i. E., Most of the dermis layers and constituents, including the papillary dermis and the reticular dermis. In other embodiments, the DMO includes primary dermal tissue, but may also include fatty tissue. In some embodiments of the present invention, DMO does not produce keratin or produces keratin in negligible amounts, thus preventing keratin cysts from forming after subcutaneous implantation in the recipient.

II. DMO sampling method and device

The DMO to be harvested can be taken from the body by any tissue harvesting means known in the art, for example, a biopsy process. The harvesting process keeps the microstructure of the harvested tissue intact. In one embodiment, the DMO can be obtained by direct biopsy and then cut to the desired size. In another embodiment, the tissue sample is obtained by direct biopsy and the biopsy yields dermal micro-organ having the desired size. In other embodiments, undesired tissue can be cut from a collected biopsy or directly taken from a micro-organ. In one embodiment, the DMO can be produced by direct biopsy and then DTMO by genetic modification of DMO in vitro. In one embodiment, the DMO or DTMO may be labeled in vitro for identification purposes, e.g., DMO or DTMO may be labeled with a chromophore, such as, for example, a chromophore that may be visible to the naked eye, Inactive, biocompatible inks or dyes containing the active ingredients of the present invention.

In some embodiments of the invention, the dermal micro-organ is harvested directly from the body. In another embodiment, the dermal micro-organ is harvested with the aid of a collection device. The internal diameter of the cutting tool used to harvest the dermal micro-organ may be, for example, about 0.5-4 mm. In other embodiments, the diameter may be, for example, 1.71 mm. In another embodiment, the diameter may be 1.21 mm. In another embodiment, the diameter may be, for example, 1-3 mm. In other embodiments, the diameter may be, for example, 2-4 mm. In one embodiment, the diameter may be, for example, 1-2 mm. In other embodiments, the diameter may be between 0.5 and 1.5 mm. In another embodiment, the diameter may be, for example, about 1.5 mm. In another embodiment, the diameter may be, for example, about 2 mm.

In some embodiments, the cutting tool has dimensions corresponding to the needle size dimensions. In one embodiment, the cutting tool is, for example, a 14GA needle. In another embodiment, the cutting tool is a 15GA needle. In another embodiment, the cutting tool is a 16GA needle. In another embodiment, the cutting tool is a 17GA needle. In another embodiment, the cutting tool is an 18GA needle. In another embodiment, the cutting tool is a 19GA needle. In one embodiment, the cutting tool is a 12GA needle. In another embodiment, the cutting tool is a 13GA needle. The wall thickness of the cutting tool corresponding to the needle size dimension may be, for example, a constant wall thickness (RW), a thin wall thickness (TW), an extra thin wall thickness (XTW) Thickness.

The tip shape of the cutting tool can also play an important role in harvesting DMO. For example, a sharp tip such as a commercially available needle may be used. As another example, the cutting tool may have a sharpened tip, e.g., by abrasion, or by using a chemical treatment agent or by electrochemical corrosion. In one embodiment, the sharp tip of the cutting tool is symmetrically shaved. The sharpening of the tip may be on the OD surface or the ID surface. For example, the tip may be sharpened by removing material from the outer or inner surface of the tip.

In some embodiments, the collected DMO may not retain the cylindrical shape after harvesting, i.e., one or more dimensions of the cross-section of the cross-section may be long and at least another dimension of the cross-section of the cross-section may be reduced. In one embodiment, for example, the at least one dimension may be 0.5-3.5 mm, and the at least one dimension may be 1.5-10 mm.

In other embodiments, the dimensions of the tissue to be harvested may be, for example, from about 5 to 100 mm in length. In other embodiments, the dimensions of the tissue to be harvested may be, for example, a length of about 10-60 mm. In other embodiments, the dimensions of the tissue to be harvested may be, for example, about 20-60 mm long. In other embodiments, the dimensions of the tissue to be harvested may be, for example, about 20-50 mm in length. In other embodiments, the dimensions of the tissue to be harvested may be, for example, 20-40 mm long. In other embodiments, the dimensions of the tissue to be harvested may be, for example, about 20-100 mm in length. In other embodiments, the dimensions of the tissue to be harvested may be, for example, about 30-100 mm in length. In other embodiments, the dimensions of the tissue to be harvested may be, for example, about 40-100 mm in length. In other embodiments, the dimensions of the tissue to be harvested may be, for example, about 50-100 mm in length. In other embodiments, the dimensions of the tissue to be harvested may be, for example, about 60-100 mm in length. In other embodiments, the dimensions of the tissue to be harvested can be, for example, about 70-100 mm in length. In other embodiments, the dimensions of the tissue to be harvested may be, for example, about 80-100 mm in length. In other embodiments, the dimensions of the tissue to be harvested may be, for example, about 90-100 mm in length. In other embodiments, the length may be about 20 mm. In other embodiments, the length may be about 30 mm. In other embodiments, the length may be about 40 mm.

If the DMO has the above dimensions, it can be maintained in the culture medium for a prolonged period of time, such as several days, a few weeks, or a few months, for example, under appropriate tissue culture conditions. For example, DMO can be maintained in vitro in designated culture media. In an exemplary embodiment, the culture medium can include growth factors, fetal bovine serum (FCS), human serum, or synthetic serum substitute (SSS). In another exemplary embodiment, the culture medium may comprise serum derived from a donor or recipient organism. In another embodiment, the culture medium may comprise autologous serum. In another embodiment, no serum is added to the medium.

In one aspect, in accordance with some embodiments of the present invention, the DTMO that is produced can be used for only a portion of the treatment period. The rest of the DTMO organization may be retrieved and maintained thereafter, and / or stored (e.g., cryogenic storage or otherwise stored) for future use. In one embodiment, the DMO is stored, e. G., Cryogenic, or otherwise stored prior to the transgenic process. In other embodiments, the DMO is stored, e. G., Cryogenic, or otherwise stored after the transgenic process.

According to one aspect of some embodiments of the present invention, as described below, a large number of dermal micro-organisms can be processed into a DTMO together in a batch process. By doing this, it is possible to process more conveniently but it will not be possible to individually determine the secretion level of each DTMO. In other embodiments, the DMO can be individually processed into DTMO as described herein.

In some exemplary embodiments of the invention, an efficacy analysis may be performed on a therapeutic agent, which may be produced and / or secreted by either a single DTMO or DTMO batch. Efficacy assays may include, for example, cell proliferation assays in which the proliferative response of the cells is predominantly dependent on the presence of the therapeutic agent in the culture medium of the cells. In one embodiment, the analysis of DTMO can utilize an ELISA assay, for example, to quantitate secretion levels of one or more secreted therapeutic agents.

In some embodiments of the present invention, the DMO collection method may be performed in such a manner that at least one or more other tissue segments of at least DMO and / or the vicinity thereof are maintained in a desired shape and / or location, , Stabilizing and supporting the skin-related tissue structure from which the DMO is harvested, extracting at least a portion of the DMO from the surrounding tissue, and separating the extracted DMO.

1 illustrates, in block diagram form, an overview of how to make and use DMO and DTMO in accordance with an exemplary embodiment of the present invention. Likewise, the DTMO can be prepared by performing the steps described separately from the bioreactor. At block 202, the DMO is taken from the object. In some embodiments of the present invention, the DMO is taken from the same subject to which future treatment will be applied. In one embodiment of the invention, the DMO is harvested from dermal tissue. Alternatively, other tissues may be harvested and used in a manner similar to that described below in connection with dermal tissue. The method described below is an example, and in some embodiments of the invention, other methods of harvesting tissue samples may also be used. If desired, the DMO or DTMO can be cryogenically stored and subsequently used (i.e. introduced into the same step in the process). As another example, in certain embodiments, the DMO can be implanted directly into a patient that has been harvested for therapeutic, cosmetic, or other physiological effects.

In order for the DMO to be a viable micro-organ, the DMO can diffuse nutrients from the nutrient medium in contact with the DMO to all the cells of the DMO, and can be one or more of a number of dimensions small enough to allow the waste to diffuse out of the DMO to the medium . By doing so, the DMO can survive in vitro for a time sufficient to selectively utilize DMO selectively as a source of therapeutic agents such as additional treatments and proteins described below. With the DMO sampling method, a DMO having an in-vitro life of several months is generally obtained.

A suitable genetic modification is prepared (block 208). Other exemplary methods of preparing the formulation include preparing a fraction of the desired amount of the modifier using a predetermined diluent, including, for example, a modifier such as a virus, and verifying the activity of the modifier do. Both of these processes are well known in the art. At this time, the DMO can be stored at a cryogenic temperature and can be injected at the same point in the future process. This can be done using a known protocol for gradual freezing of tissues and cells, for example DMEM medium containing 10% DMSO.

At block 210, the DMO is transgenic. As described above, various gene modification methods are known and can be used in connection with the present invention. As an example, the following description is based on the insertion of a gene into cells of DMO using a viral vector. This process is well known and will not be described further except for the specific methods and devices for introducing the virus into the DMO.

At block 212, the production rate and secretion rate of the therapeutic agent are selectively tested for genetically modified DTMO. There are various methods for measuring the secretion amount, for example, ELISA, other immunoassays, and spectral analysis methods. In addition to the secretion amount, for example, the sterility and activity of the secreted protein is selectively tested. These tests can be performed periodically or continuously on-line. At this time, the DTMO can be stored at a cryogenic temperature for future use.

At block 214 and block 216, the amount of DTMO required to achieve the desired therapeutic effect is determined. As discussed below, the dosage required for treatment may be determined from population statistics on secretion rates, patient parameters, and anticipated or known correlations between in vitro secretion levels and in vivo serum levels.

At block 218, the desired number of DTMOs is selected. Load the DTMO into the transplant tool. An example of a portable device is described below.

If the DTMO needs to be moved before it is transferred to the implant, it optionally maintains temperature, humidity, etc. under maintenance station or maintenance conditions (220), maintaining the level of viability while the DTMO is carried. The remaining DTMOs are optionally maintained in vitro for future use. It can be at low temperature incubator conditions (4 캜), under the conditions as described above, under warm incubator conditions (30 - 37 캜), and can extend its viability in vitro or under cryogenic conditions have.

At block 224, the DTMO subset is ported to the entity. Exemplary implementations of the implantation method are described below. Other methods for this will be known to those skilled in the art and are primarily determined by the specific geometry of the micro-organs used. In animal studies, DMO or DTMO is known to be viable in vivo, in the sense that DTMO continues to produce and secrete therapeutic agents for several weeks and months after transplantation. In animal experiments, the therapeutic amount is produced for a period of 160 days or less (or more). The tissues of the DMO or DTMO appear to be integrated or appropriately housed in the tissue of the individual to which they are implanted (in particular, when the tissue is implanted into a tissue of the same species as the tissue from which the tissue is harvested), the cells contained in the DMO or DTMO are treated Produce and secrete the formulation continuously.

The in vivo efficacy of DTMO is selectively confirmed (block 228). For example, based on this assessment, and / or based on past patient data (block 226), administration to a patient by increasing the amount of implanted material, or removing some of the implanted material, as described below The dose can be adjusted (block 230). If the efficacy of the transplant varies, DTMO may be further transplanted.

Genetic modification can typically involve genetic manipulation of selected genes or genes into cells that cause the cells to produce and selectively secrete a desired therapeutic agent such as a protein. In one embodiment of the invention, at least part of the process of maintaining the DMO during the transgenic process, as well as transgenic transformation itself, can be carried out in the bioreactor.

In some exemplary implementations of the present invention, a method of harvesting DMO from an individual may be performed in a desired configuration and position, such as, for example, in a targeted harvesting section And / or maintaining a skin-related tissue structure associated with the DMO, which is generally located in the skin. For example, by attaching more than a certain portion of the epidermal portion to a predetermined surface so that more than a portion of the skin-associated tissue structure can be lifted to maintain the desired shape and / or position, The epidermis can be lifted. In some exemplary embodiments, the process of attaching the epidermis to a predetermined surface can include applying a vacuum condition, for example, as described below. As another example or additionally, the process of attaching the epidermis to a predetermined surface may include applying an adhesive to the surface.

Turning now to FIG. 2, which schematically illustrates a flowchart of a DMO harvesting method in accordance with some illustrative embodiments of the present invention and FIG. 3A, which schematically illustrates an example of a step of harvesting a DMO 3024 located within a skin- -3G.

As shown in block 2002, the method may optionally include topical administration of an anesthetic, for example, around the DMO to be anesthetized, as is known in the art.

The use of DTMO in protein or RNA-based therapies may, in certain cases, require the use of multiple DTMOs. As described elsewhere, DMO and DTMO can be kept in vitro for long periods or stored at cryogenic temperatures for future use. That is, in some cases, a plurality of DMOs may be continuously collected from the same individual during a single treatment period. In this way, an individual can harvest multiple DMOs from an individual for future use, without having to perform separate individual harvesting operations for each individual DMO for several days. In one embodiment, a collector marker template can be used (step 2004), prior to placing the harvesting device on the surface of the individual's skin, to mark several sites to be harvested. In one embodiment, the collector marker template is placed on the skin surface of an individual and then marked on the surface of the skin, for example to indicate areas for application of local anesthesia, alignment lines and harvest lines. In one embodiment, the surface is marked with a surgical pen or marker. In one embodiment, the surface is marked using a non-permanent dye or ink.

As shown in block 2004, the method includes attaching a support structure (e.g., Figure 4C; Figure 5A; Figure 6; Figure 11) to a predetermined harvesting portion such that the support structure, or a portion thereof, And / or < / RTI > In some embodiments, the contact between the support structure of the present invention and the surface of the skin of the subject must be air-tight such that a vacuum seal can be formed at a later stage. In one embodiment, the harvesting unit is an individual or the like. In another embodiment, the harvesting portion is the abdomen of the individual. In another embodiment, the harvesting portion may be another location of the individual's body.

As shown in block 2006 and FIG. 3A, a support structure (e.g., FIG. 4C; FIG. 13A; FIG. 6), which may include a vacuum chamber (FIG. 11 1130) and a guide channel 2008 (FIG. 11 1108). 11) can be used under vacuum to stabilize and support skin related tissue structures, including epidermis 3000, dermis 3002, and fat 3004 tissue layers, for proper extraction of DMO. For example, application of a vacuum condition may cause a vacuum to form within the vacuum chamber, causing the skin surface of the skin-related structure to be drawn into the support structure, wherein the center channel is in contact with the skin surface layer of the skin- It can support the dermal layer.

Herein, the term "guide channel" may also be referred to herein as a "needle guide channel ".

Under vacuum, the central channel can provide support for skin-related tissue structures to take the form that the dermal tissue is in the center channel. In some exemplary embodiments, the vacuum chamber includes a raised protrusion. In other exemplary embodiments, the vacuum chamber includes two raised protrusions. When a support structure comprising one or two raised protrusions is used, the raised protrusions may additionally support a skin epidermal layer and a dermal layer of the skin-associated structure.

In certain cases, with the application of a vacuum using a vacuum chamber with two raised protrusions, the precise geometry of the skin-related structure is formed, so that the dermal tissue may be harvested and the plug of the epidermal tissue may not be harvested.

In the exemplary embodiments, the vacuum condition may cause the skin-related structure to remain on the inner support surface of the vacuum chamber, if present, in the central channel and the raised protrusion. In one embodiment, the guide channel 3008, which may be tubular in shape, may provide guidance and / or stability for inserting and / or using a cutting tool so that correct cutting along the cutting axis can be made. In some embodiments, the cutting axis is coaxial with the guide channel. While the coring tube is coaxial with the guide channel, the coring needle may not always be located in the vertical center of the center channel. In one embodiment, the core ring tube is located at the horizontal center of the center channel.

In certain embodiments, the internal dimensions of the support structure, including the vacuum chamber, may be 3.0 - 8.0 mm. In one embodiment, the dimensions may be, for example, 3.0 mm in diameter. In other embodiments, the dimensions may be, for example, 3.5 mm in diameter. In another embodiment, the dimension may be, for example, 4.0 mm in diameter. In other embodiments, the dimensions may be, for example, 4.5 mm in diameter. In other embodiments, the dimensions may be, for example, 5.0 mm in diameter. In another embodiment, the dimension may be, for example, 5.5 mm in diameter. In another embodiment, the dimensions may be, for example, 6.0 mm in diameter. In other embodiments, the dimensions may be, for example, 6.5 mm in diameter. In other embodiments, the dimensions may be, for example, 7.0 mm in diameter. In another embodiment, the dimension may be a diameter of, for example, 7.5 mm. In other embodiments, the dimensions may be, for example, 8.0 mm in diameter.

In one embodiment, a suitably sized support structure with a vacuum chamber of a particular inner diameter is predetermined prior to actual harvesting.

As shown in block 2008 and in Figure 3B, a support structure 3006 is formed using a introducer (e.g., Figure 4D), including, for example, the inner needle 3010 and the outer sleeve 3012; The tissue associated with the skin can be perforated by inserting it into the skin-related tissue structure at the penetration point through the introducer into the guide channel of the skin. This single perforation site can be used for any additional inflow and outflow into and out of the skin-related structure. In this way, damage to the individual and scarring are limited. The introduction of the cutting tool through the outer sleeve of the introducer prevents or minimizes the extraction of the epidermal tissue.

The introducer may comprise, for example, a tubular member, an inner needle and an outer sleeve. In some embodiments, the outer sleeve is fitted with an inner needle in it. They can be incorporated and inserted into skin-related structures. In certain embodiments using a support structure having a vacuum chamber that includes a raised proximal projection, the skin-associated structure is arranged such that the introducer is inserted at a point perpendicular to the surface of the skin as a whole at the point of penetration. In one embodiment using a support structure with a vacuum chamber comprising a ridge-shaped proximal projection, the tip of the inner needle reaches the tissue region of the first zone, i.e., the ridge-shaped prominent protrusion 3007 region, The introducer can be inserted into the skin-related structure such that the fabric part reaches approximately midway below the first raised protrusion. In one embodiment, after inserting the introducer, the distal end of the outer sleeve is pierced through the entire layer of skin in the perforation and is located in the basal fat layer.

If the step of the block 2008 using the introducer to puncture the skin is omitted, the skin full-layer plug will be harvested prior to harvesting the dermal micro-organ in the step of the lower block 2012, Additional processing of the tissue to remove the plug may be necessary.

In one embodiment, the inner needle is beveled. In another embodiment, the inner needle is not a slope. In one embodiment, the introducer inserts the bevel of the inner needle downward. In another embodiment, the introducer inserts the slope of the inner needle upward. In another embodiment, the introducer inserts a sloped surface of the inner needle at an intermediate angle between upward and downward.

The following exemplary embodiment (block 2010) relates to the use of a support structure, including a vacuum chamber including at least proximal bulging protrusions 3007 and distal bulging protrusions 3009.

As shown in block 2010 and in Figure 3C, the outer sleeve 3012 of the introducer passes the introducer through the inner guide channel and through the skin layer 3000 and dermal layer 3004 of the skin- By inserting the fat layer 3002, which is located in the lower layer 3002. After insertion of the introducer, the inner needle is withdrawn from the skin-related structure and the outer sleeve is placed in a position where the distal end of the outer sleeve remains approximately midway below the prominent protrusion. With this operation, the outer sleeve is coaxially positioned with the inner guide channel, which can also be the cutting axis, and the tip of the outer sleeve reaches the fat 3002 of the skin-related tissue structure supported by the proximal projection 3007 .

The outer sleeve 3012 can be constructed of a thin tube, hollow rod, or any other suitable thin, normally straight, material that can penetrate the required skin layers and be disposed around the inner needle. In one embodiment, the outer sleeve may have an inner diameter corresponding to a needle size of 12-19 GA, such as about 14 GA. In one embodiment, the outer sleeve can include an appropriately sized plastic tube. In one embodiment, the outer sleeve comprises a high density polyethylene (HDPE) tube. In another embodiment, the outer sleeve comprises a polytetrafluoroethylene (PTFE) tube. In another embodiment, the outer sleeve comprises a fluorinated ethylene propylene (FEP) tube. In one embodiment, the length of the outer sleeve is about 10-100 mm. In one embodiment, the length of the outer sleeve is about 40 mm. In one embodiment, the outer sleeve is inserted into the skin-related structure through about 5-20% of the length through the perforation. In one embodiment, the outer sleeve is inserted into the skin-related structure through about 10-15% of the length through the perforation. In one embodiment, the outer sleeve is inserted into the skin-related structure through about 12.5% of the length through the perforation.

The above-described embodiments describe that the outer sleeve penetrates into the fat layer. In other embodiments, the outer sleeve may be inserted into the dermal tissue. In another embodiment, the outer sleeve can be inserted into the subcutaneous space.

The length through which the outer sleeve 3012 penetrates the skin through the skin into the dermis or fat, or subcutaneous space, is typically 1 to 15 mm, or, in one embodiment, about 5 mm. For example, the outer sleeve can be manually inserted as part of the introducer and guided to a desired depth into the dermis, fat, or subcutaneous tissue beneath the proximal protrusion. The outer sleeve is then coaxial with the cutting axis in the center channel, thereby allowing the dermis to be harvested by the cutting tool.

In one embodiment, the use of an outer sleeve prevents the puncture opening from being exposed to rotational and forward motion of the cutting tool, thereby preventing additional skin trauma at the mouth.

As shown in block 2012 and Figures 3D and 3E, DMO can now be cleaved from skin-related tissue. The cutting tool 3014 can be coaxially inserted into the guide channel 3008 and the outer sleeve 3012 and the outer sleeve guides the cutting tool along the cutting axis (Figs. 5A and 6) to the skin related structure. In one embodiment, the guidance of the cutting tool through the outer sleeve allows the cutting tool to directly reach the dermal tissue 3004 of the skin-related structure (3E). As an aid for cutting, the cutting tool can rotate as it advances the tube toward the distal end of the device. The motor can be used to rotate the cutting tool. The motor may be, for example, a pneumatic motor drill or an electric motor drill. In some embodiments, a medical drill may be used to rotate the cutting tool. In one embodiment, a drill collet 3016 is attached to one end of the cutting tool as is known in the art.

In one embodiment, the method may include rotating the cutting tool while advancing the cutting tool, e.g., toward the distal end of the support structure. For example, the dermal tissue for DMO can be cut more smoothly by manually or automatically rotating the coring tube 3014 using a medical drill or other suitable tool or rotating mechanism. For example, a medical drill can be connected using a drill collect 3016 at the proximal end of the coring tube. An example of a medical drill is a drill in Germany, which may include control units, motors, connection cords, handpieces and / or foot switches with catalog numbers GD650, GD658, GB661, GB166 and GB660, D-78532 Tuttlingen, Am Aesculap AG & Co. located in Aesculap Platz. Aesculap Microspeed Drill from KG; Nouvag medical drill, TCM-3000-BL, and handpiece (catalog numbers 3285 and 1710, respectively). Using such a drill or any other suitable drilling or rotating mechanism it is possible to apply a suitable rotational speed, for example a relatively high rotational speed, for example greater than 1,000 RPM, for example 1,000 RPM to 10,000 RPM, , The cutting edge of the cutting tool can be rotated. For example, the tube 3014 may be rotated at a speed greater than 2,000 RPM, for example, at a speed of about 7,000 RPM. In other embodiments, it may be rotated at a relatively low rotational speed of less than 1,000 RPM, or may not be rotated at all as described below. Alternatively, the rotational speed of the drill may vary depending on the vibration mode, i.e., the rotational direction may be periodically changed between "clockwise" and "counterclockwise ". The coring tube can be advanced manually or automatically, e.g., toward the distal end of the support structure, e.g., toward the distal raised protrusion 3009, while being rotated by the drill.

In one embodiment, a method of cutting a dermal micro-organ can include stopping the advancing movement of the coring tube at a specific location. In one embodiment, when the drill collect 3016 abuts the proximal end of the introducer sleeve 3012 at the outer surface of the support structure 3006, it acts as a hard stop to prevent further advancement of the coring tube . In another embodiment, when a member mounted on the outer distal portion of the coring tube, such as a cap (FIG. 13) 1302 wrapping the coring tube, is brought into contact with the outer surface of the support structure, So that the coring tube can be prevented from continuing to advance.

In certain exemplary embodiments, a method of cutting a DMO using a support structure having a vacuum chamber having at least one distal bulging protrusion may include moving the advancing motion of the coring tube against a tip 3E may advance and remain in the area of the skin-related structure located below the distal protruding portion 3009. [ In one embodiment, the distal tip of the coring tube may be located in the fat layer when the drill collector abuts the introducer sleeve at the outer surface of the support structure. In one embodiment, when the drill collector abuts the introducer sleeve at the outer surface of the support structure, the tip of the coring tube may be located below the distal protruding portion and into the lipid layer.

In one embodiment, the geometric arrangement of the skin-related structure formed in the vacuum chamber having at least the distal bulging protrusions ensures that the distal tip of the coring tube passes through the dermal / lipid interface at the end of the advancing movement, Lt; / RTI > The dermis / fat border is weakly binding. In one embodiment, the weak binding between the dermis and fat may allow the separation of dermal tissue samples from body fat during DMO harvest.

The cutting tool may comprise any suitable cutting tool, for example a coring tube (e.g., FIG. 4E; 4010). The coring tube may comprise a generally symmetrically scored tubular tool, e.g., a subcutaneous tube that is sharpened to the desired shape of the cutting edge. The coring tube may include, for example, a standard medical grade tube having a thin wall portion, e.g., a wall thickness of 0.05 mm - 0.3 mm. The coring tube may have an inner diameter of, for example, 0.5 mm to 4 mm. In one embodiment, the inner diameter may be 1-2 mm. In other embodiments, the inner diameter may be 1-3 mm. In another embodiment, the inner diameter may be 2-4 mm. In another embodiment, the inner diameter may be 0.5-1.5 mm. In one embodiment, the inner diameter may be about 1.21 mm. In other embodiments, the inner diameter may be about 1.5 mm. In another embodiment, the inner diameter may be about 1.71 mm. In another embodiment, the inner diameter may be about 2 mm. In one embodiment, the core ring tube has a needle having a dimension of about 14 GA. In another embodiment, the coring tube comprises a needle having dimensions of about 12 GA, 13 GA, 15 GA, 16 GA, 17 GA, 18 GA, or 19 GA. The dimensions of the coring tube, e.g., the diameter and / or dimensions of the introducer, may be predetermined depending on the volume and / or size of the DMO to be harvested. The coring tube may have a sharpened end ("tip") designed to be used as a cutting edge. In one embodiment, the sharpened edge is cut at the outer diameter. In another embodiment, the tip is trimmed in the bore. The coring tube may be inserted through the outer sleeve into the tissue structure associated with the skin, in order to prevent epidermal tissue from being harvested. In one embodiment, using a support structure having at least proximal bulge-like protrusions, a precise geometric arrangement is established in the skin-related structure under vacuum to ensure that the epidermal layer and / or the epidermal plug is not harvested.

In some exemplary embodiments of the present invention, as will be described below, in order to facilitate separation of the harvested tissue from the inner surface of the cutting tool during subsequent operations, and / or to reduce any force exerted on the tissue upon cleavage , the inner surface and / or outer surface friction is at least a portion of the tube may also be coated with a low material, e.g., Teflon ^®, parallel Lin (Parylene), or other suitable coating material.

As shown in block 2014 and Figs. 3F and 3G, the harvesting method includes the recovery of DMO 3024. After the coring tube is advanced to the hard stop and the distal tip of the coring tube is positioned in the skin related tissue structure located below the distal raised protrusion (e.g., FIG. 6), the drill collect 3016 is opened (FIG. 4H) Remove the drill while keeping the core ring tube in position. In one embodiment, an outer cap, such as a luercap, may be used to maintain the position of the core ring tube. The collect 3020 / partition 3021 of the syringe 3022 assembly is then attached to the coring tube (Figures 4B, 4A 4004 and 4 4012, respectively).

In one embodiment, male luer cap 1302 is affixed to the outer surface of the coring needle 1304, e.g., glued. The clamping force to pull out the drill and fix the glued cap to the position of the syringe during the connection process ensures that the coring needle does not move even during the final sampling stage of the DMO. This can prevent the possibility of DMO loss.

First, a collector 3020 is attached. This is a stand-alone collet, not a medical drill. The collector can slide over the coring tube when in an open position. When the collector is in place and mounted in the closed position, the partition 3021 is pushed over the needle and is engaged with the collector. In one embodiment, the partitions include a needleless valve. In one embodiment, the needleless valve provides an airtight seal. The collect and the bulkhead are collectively called a needleless valve assembly. The collector prevents the bulkhead from pushing the coring tube in the forward direction while attaching the bulkhead. The syringe 3022 may be coupled to the needleless valve assembly after the septum is pierced with a coring tube (e.g., by a screw) and coupled with the collector. Pulling the syringe flapper creates a vacuum in the coring tube. At this time, the entire assembly with the coring tube can be withdrawn (FIGS. 3F and 3G), which causes the cut tissue to be drawn into the syringe.

In one embodiment, the vacuum condition is applied at the same time as the coring tube is removed from the skin-related tissue structure and the DMO is collected, for example, into the syringe body. In another embodiment, the coring tube is removed from the skin-related tissue structure and then a vacuum is applied to the coring tube to allow the DMO to be recovered, for example, to the syringe body. In another embodiment, the vacuum state is applied to the coring tube while the coring tube is present in the skin-associated tissue structure and the DMO is recovered, for example, to the syringe body. In another embodiment, the drill is detached from the coring needle 1304, with the glued cap 1302 held in place (Figure 13). Then, the armure syringe 1306 is directly connected to the glue-capped cap. When pulling the syringe plunger and removing the syringe / coring needle assembly from the skin tissue structure, the DMO is aspirated directly into the syringe body. The combination of the coring tube and syringe with the luer cap eliminates the need for a collect and needleless valve assembly.

In one embodiment, the syringe is partially filled with saline or other suitable liquid so that the tissue sample is recovered to a fluid environment that supports tissue viability.

The partition 3021 is necessary to ensure an air-tight connection between the syringe 3022 and the inner space of the coring needle. If the syringe is directly coupled to the collector without a septum, a vacuum will not form on the coring tube because the collector is not normally airtight when the syringe plunger is pulled. 4A 4012 illustrates an example of a bulkhead that may be used in a method of harvesting the DMO of the present invention in certain embodiments.

In another embodiment, the recovery of DMO is accomplished by evacuating the cutting tool in the skin-related structure, wherein the DMO is retained in the cutting tool. The DMO 3024 can then be withdrawn from the cutting tool using positive pressure, for example, by coupling the syringe to the proximal end of the cutting tool and applying positive pressure with the syringe plunger, . In addition, an appropriate fluid, such as a sterile fluid, can be used to assist in the recovery of DMO from the cutting tool 3014.

In another embodiment, the DMO can be carefully removed from the cutting tool using mechanical means, such as, for example, tweezers or similar tools, and the DMO can be removed using the means described above, I can catch it.

As shown in block 2016, the device can then be evacuated from the harvesting station. At this time, the outer sleeve can be manually removed from the skin-related tissue.

Those skilled in the art will appreciate that any combination of the above operations may be implemented to perform harvesting in accordance with embodiments of the present invention. Furthermore, other operations or series of operations may be used.

4C-E which illustrate an exemplary embodiment of a harvesting apparatus, Fig. 4C shows an example of a support structure with a vacuum source attached thereto; Fig. 4D shows an example of a introducer, an inner guide needle 4006 and an outer sleeve (4008-white tube); 4E (4010) is an example of a coring tube; 4014 is an example of a medical drill that can be used to rotate a cutting tool, and 4012 is an example of a drill collet for attaching to a coring tube. Figures 4A-B illustrate examples of syringes (Figure 4A 4012), partitions (Figure 4A 4004), and syringe collect (Figure 4B) for use in recovering dermal micro-organ.

According to some embodiments of the present invention, the manual process described above can be easily performed by an integrated device (unaided) designed to perform some or all of the above-described processes for collecting DMO. For example, in one implementation of the harvesting method, the integrated apparatus includes a positioning and insertion guide (FIG. 4D) of the introducer, an insertion guide (FIG. 4E 4010) of the coring tube, Motion control, and / or DMO recovery from the coring tube. Such a device can perform a relatively simple operation when performing the harvesting process.

5A, which schematically illustrates a dermis harvesting apparatus 5000 according to another exemplary embodiment of the present invention. As used herein, the term "support structure" refers to the body of the device used to support the skin-associated tissue structure. The term "support structure" may also be referred to herein as an "apparatus ". In this hack, the term "device" has all the properties and properties of a "support structure. &Quot;

The apparatus 5000 may include a vacuum chamber 5001 including a guide channel 5003 and a raised protrusion 5006. The raised protrusions 5006 may have a predetermined size and / or thickness, for example, designed to form a " high profile "consisting of a skin tissue monolayer of flattened orientation generally raised above the trajectory of the coring tube 5016. [ And the like. For example, section 5006 is higher than other sections of chamber 5016 so that fat layer 5018 can be pulled up to section 5006 and supported along the orbit of coring tube 5016. As a result, after collecting the DMO of a predetermined length, the coring tube 5016 advances to the fat layer 5018, thus separating the DMO from the tissue surrounding the DMO. The collected DMO may be in it while removing the coring tube 5016 from the body, or may apply a vacuum to the distal end of the coring tube to suck DMO from the coring tube. Due to the configuration of the device 5000, it may not be necessary to make a "drain" incision in the skin, and thus the DMO can be recovered with only one incision.

In some exemplary implementations of the present invention, the apparatus 5000 also includes a means for directing the coring tube 5016 to a predetermined distance along the chamber 5001, such as, for example, 5018. The drill stopper 5008 is capable of manually advancing to a position where it has reached the position where it has reached the position where it has reached the position where it reaches the position

In some exemplary embodiments of the present invention, the apparatus 5000 may also include a vacuum chamber 5001 and a vacuum conduit 5004 coupled to the vacuum source 5002.

5B, which schematically illustrates a cross-sectional view of a harvesting apparatus 5050, which is now embodied for externally supporting a skin-related tissue structure at a desired location, according to another exemplary embodiment of the present invention.

In some embodiments, the apparatus 5050 can include two channels 5064 that are positioned over a portion of the chamber 5001 along both sides of the chamber 5001, so as to clamp the skin layer 5020. The channel 5064 can be located at a desired height, for example, at a height approximately equal to the height at which the center of the DMO is taken, e.g., centered. In one embodiment, the center channel is located about 2 mm lower than the top surface of the vacuum chamber, so that clamping is stable and / or can support tissue cleavage. In this exemplary embodiment, the apparatus 5050 further includes an inner surface 5060 of the channel 5064, such as an inner surface or an outer surface of the channel 5064, so as to allow the tissue to be clamped outside without substantially affecting the vacuum condition applied to the chamber 5001. [ Two flexible membrane members 5058 can be included on either side of the surface. In another embodiment of the present invention, apparatus 5050 may not include member 5058 and / or channel 5064.

According to the exemplary embodiment of Fig. 5B, by clamping the skin-related tissue structure supported in the vacuum chamber outward, it is possible to improve the stabilization of the dermis 5014 and / or the retention of fat 5018 into the vacuum chamber 5001 . For example, the clamping tool 5070 can be used to "pick" the skin-related tissue structure, e.g., symmetrically, supported in the vacuum chamber 5001. Both clamping ends 5054 of the clamping tool 5070 can be inserted into the channel 5056, respectively. The tool 5070 can be closed so that the clamping end 5054 can press the flexible member 5058. [ That is, the skin-related tissue structure in the chamber 5001 can be clamped on the side without substantially affecting the vacuum state in the chamber 5001. The clamping force exerted by the clamping end 5054 may correspond to a constant or variable force of, for example, the spring 5062 or other suitable mechanism.

6 and 11, which schematically illustrate implementations of the harvesting apparatus according to some exemplary implementations of the present invention.

The apparatus 6000 includes a guide channel 6003 (Figures 11 and 1108), two raised protrusions 6007 (proximal) and 6006 (distal)) (Figure 11, 1107 and 1109, respectively) and two raised protrusions And a vacuum chamber 6001 (Figs. 11 and 2030) with a central channel of the vacuum chamber 6001. The apparatus 6000 may further include one or more vacuum channels and a vacuum conduit 6004 (FIG. 11, 1126) to fluidically couple the vacuum chamber with one or more vacuum sources 6002. The raised protrusions 6006 and 6007 are designed to be able to form, for example, a "flattened surface" composed of a skin tissue monolayer of generally flat orientation on the orbit of the coring tube 6016, And / or < / RTI > The raised protrusions 6006 and 6007 may or may not have the same size and shape. For example, sections 6006 and 6007 may be higher than other sections of the chamber so that epidermal layer 6020, dermal layer 6060 and fat layer 6018 can be raised respectively to sections 6006 and 6007 , So that in some embodiments, the dermal tissue layer 6060 is secured within a central channel within the orbit of the coring tube 6016. [

Applying a vacuum to a device with a vacuum chamber containing two raised protrusions builds up a skin-related tissue structure in a precise geometrical arrangement, for example, so that the dermis is harvested and the epidermal layer of the skin is not harvested. In the presence of proximal bulgeous protrusions along with the use of the introducer, the plug of epidermal tissue is prevented from being collected at the proximal end of the DMO. After picking up the DMO to a predetermined length, the coring tube 6016 can be advanced to the fat layer 6018 to separate the DMO taken from the tissue surrounding the DMO. The collected DMO may remain in the coring tube 6016 during removal from the body, or may apply a vacuum to the proximal end of the coring tube to inhale the DMO from the coring tube.

Due to the configuration of the device 6000, it may not be necessary to make a "drain" incision in the skin, and thus the DMO can be recovered with only one incision.

In some embodiments of the present invention, the internal width of the vacuum chamber in apparatus 5000 and / or 6000 is about 3.5 mm. In one embodiment, the center channel may have a width of, for example, about 4 mm. In other embodiments, the center channel may have a width of, for example, about 3.0 mm. Further, in some embodiments, the center channel may have a height of, for example, about 5 mm, excluding the protrusions. In other implementations, other ranges for the width and / or height of the center channel may be used, such as 3-25 mm, and in some embodiments of the invention, for example, any titration in the range of 3-25 mm Dimensions may be used. The length of the central channel may be generally similar to the length of the DMO being harvested, for example about 30 mm in length; For example, in the range of 5-100 mm. In other embodiments, the dimensions of the channel length may be, for example, about 10-60 mm in length. In other embodiments, the dimension of the channel length may be, for example, about 20-60 mm in length. In other embodiments, the dimensions of the channel length may be, for example, about 20-50 mm in length. In other embodiments, the dimensions of the channel length may be, for example, about 20-40 mm in length. In other embodiments, the dimension of the channel length may be, for example, about 20-100 mm in length. In other embodiments, the dimensions of the channel length may be, for example, about 30-100 mm in length. In other embodiments, the dimensions of the channel length may be, for example, about 40-100 mm in length. In other embodiments, the dimension of the channel length may be, for example, about 50-100 mm in length. In other embodiments, the dimensions of the channel length may be, for example, about 60-100 mm in length. In other implementations, the dimensions of the channel length may be, for example, about 70-100 mm in length. In other embodiments, the dimensions of the channel length may be, for example, about 80-100 mm in length. In other embodiments, the dimensions of the channel length may be, for example, about 90-100 mm in length. In other embodiments, the length may be approximately 20 mm. In other embodiments, the length may be about 30 mm. In other embodiments, the length may be about 40 mm.

Before the DMO is actually harvested, the inner needle and outer sleeve (Figure 4D, 4008) may be drilled to puncture the skin so that the epidermal tissue is not harvested and to insert a portion of the outer sleeve through the skin from the perforation into the fatty tissue, The device 6000 can be used. This introducer can be inserted into the guide channel 6003 of the device 6000 where the tip of the outer sleeve reaches the area of the proximal bulge 606 and the tip of the needle passes over the distal end of the outer sleeve . In one embodiment, the tip of the outer sleeve penetrates through the skin layers to the fat layer. In one embodiment, the inner needle is a 14GA needle. In another embodiment, the inner needle is a 12GA needle. In another embodiment, the inner needle is a 13GA needle. In another embodiment, the inner needle is a 15GA needle. In another embodiment, the inner needle is a 16GA needle. In another embodiment, the inner needle is a 17GA needle. In another embodiment, the inner needle is an 18GA needle. In another embodiment, the inner needle is a 19GA needle.

When the inner needle is withdrawn, the outer sleeve remains and may extend into the dermal tissue 6060 in the guide channel 6003. As another example, the outer sleeve may be left and extend into the fat tissue 6018 in the guide channel 6003. In one embodiment, the outer sleeve may be roughly in the mid-point of the proximal raised protrusion 6007 at the insert. The outer sleeve can thereby serve as a "sleeve" that can introduce the coring needle directly up to the fat and just before the dermal tissue to be harvested. Thus, the outer sleeve prevents the epidermal tissue from being harvested. In other embodiments, the outer sleeve may serve as a "sleeve" through which the coring needle can be introduced directly to the dermis to be harvested. In addition, the outer sleeve protects the perforations from rotation and forward motion of the coring needle to prevent further trauma to the perforations. In certain cases, the outer sleeve may have a low coefficient of friction to prevent resistance to rotation and forward motion of the coring needle, thus avoiding unwanted heat generation.

In some exemplary embodiments of the present invention, the apparatus 6000 also includes a core ring 6016 with a predetermined distance along the chamber 6001, for example, a coring tube 6016, May include a drill collect 6008 or glued cap to act as a hard stop capable of manually advancing to a location that has reached the adipose tissue 6018 within the protrusion 6006. [

Tool 6016 may be used to rotate tool 6016 at a rotational speed suitable for cutting dermal tissue, for example, at a relatively high rotational speed, e.g., greater than 1,000 RPM, e.g., from 1,000 RPM to 10,000 RPM , And may be connected to the motor, as described above. The cutting edge of the cutting tool can be rotated. For example, the tool 6016 may rotate at a speed greater than 2,000 RPM, for example, at a speed of about 7,000 RPM. Once complete, the forward and rotational movement of the tool 6016 may be stopped and the cutting tool 6016 may be withdrawn with the DMO collected therein, i.e., the cutting tool may be removed from the collection section. The DMO can be used to push the DMO out of the back end (not shown) of the cutting tool 6016, for example, using a syringe to flush the tool with, for example, aseptic fluid, Can be removed from the cutting tool 6016 using a vacuum source for evacuation.

One of ordinary skill in the art will appreciate that the DMO can be harvested by forming a single incision or puncture site in the device 6000. Moreover, the device 6000 can effectively utilize a relatively thick skin area, e. G., For harvesting DMO from the back region of the donor.

In some embodiments, members of the harvesting device may be disposable.

10A and 10B illustrating embodiments of a syringe equipped with a partition and a collector. 10A shows the syringe coupled with a partition and a collector at the rear end of a cutting tool inserted into the guide channel of the support structure and the outer sleeve, wherein the support structure is connected to a vacuum source. Fig. 10B shows the coupling of a collet with a needle and a needleless valve at the rear end of the coring needle.

10A and 10B illustrate an embodiment of the present invention wherein the arm syringe is coupled to the rear end of the coring needle via a collect and needleless valve. FIGS. 10A and 10B illustrate implementations that may be used to draw the DMO out of the cutting tool, i. E. Into the syringe body.

It will be appreciated by those skilled in the art that the above-described harvesting methods and / or devices, for example, according to embodiments of the present invention may include introducing a thin tissue cutting instrument into the dermis. Thus, the sampling method and / or apparatus according to embodiments of the present invention provide a minimally invasive method of harvesting a desired tissue, wherein the DMO can be harvested while minimally damaging the outer skin surface .

While some embodiments of the invention described herein may refer to a DMO collection method and / or apparatus, those skilled in the art will appreciate that, in accordance with other embodiments of the present invention, one of the methods and / At least some of which may be implemented for any other procedure, such as any other procedure, such as plastic surgery, skin surgery or tissue harvesting. For example, a method and / or apparatus in accordance with embodiments of the present invention may be used to harvest dermal tissue for use as a filler, for example, in subsequent implants.

In some embodiments of the invention, systems and methods are provided for handling or processing in vitro ("in vitro") dermal microcirculation. In some embodiments, the dermis MO may be placed directly in the tissue culture well or the transduction chamber for further processing. In some embodiments, for example, if the DMO is present in the coring tube when it is removed from the skin, the DMO can be removed from the coring ring by using a biologically compatible fluid, such as saline or a culture medium, It can be flushed out of the tube. Through flushing the DMO, it can be flushed directly into the chamber of the bioreactor. As another example, a vacuum may be applied to the back end of the coring tube to directly "push " the DMO, for example, directly into the chamber of the bioreactor.

II. DMO / DTMO transplantation methods and devices

In some embodiments of the present invention, systems and methods for transplanting DTMO are provided. After manufacturing and / or processing the DMO by making and / or processing, e.g., by genetically modifying the DMO, the modified DMO or DTMO may be transplanted to the patient again, e.g., for protein or RNA based therapy. The number of total or partial DTMO transplanted can be determined by the desired therapeutic capacity of the secreted protein. DTMO can be implanted in subcutaneous, dermal tissue or other locations in the body. Subcutaneous transplantation using a transplantation needle can, for example, cause DTMO to remain in a linear form in the subcutaneous space. Transplantation in a linear form may be helpful in facilitating location, for example, to reduce the dose of the therapeutic protein or discontinue treatment, if the DTMO needs to be removed later or by in-situ removal . Other known geometric implant patterns may be used. In addition, linear transplantation can also help integrate dermal tissue into the surrounding tissue.

7, 8A-E, and 9A-E will be described below. Figure 7 schematically depicts a flow chart for a DMO / DTMO implantation method according to some illustrative embodiments of the present invention, and Figures 8A-E present some implementations for the members of the implantation device. The transplantation methods described herein refer to transplantation of DMO or DTMO, and the terms may be used interchangeably with the description of transplantation methods and devices. For ease of reading, the DTMO is referred to in the following description where the term "DMO" is perceived as being interchangeable with the term "DTMO" in the following description.

As shown in block 7002, the DTMO may be loaded into the syringe (Fig. 8A). For example, the DTMO can be inhaled into the loading syringe. Loading may involve drawing up a biologically compatible fluid, such as saline or culture medium. After DMTO is loaded in the syringe, a transplantation tool, such as a transplant needle (Figure 8B), may be attached to the syringe.

As shown in block 7004, in some exemplary embodiments of the present invention, the DTMO optionally connects the loading syringe with the DTMO, with the surrounding sterile saline fluid, to the proximal end of the implant needle (Figure 8B) , The DTMO can be carefully loaded with a needle using a positive pressure to be loaded into a transplantation tool. As another example, the DTMO can be sucked directly into the transplantation needle through the distal portion of the transplantation needle, for example, by pulling on the plunger of the syringe attached to the proximal end of the transplantation needle. Optionally, the tip of the needle may have a removable short silicone tube extension or the like secured thereto to facilitate suction of the DTMO through the distal portion into the needle cannula, while pulling the plunger of the syringe.

The implantable needle may have any suitable diameter, for example, 17 GA-8 GA diameters. In one embodiment, the diameter of the implant is about 10 GA diameter. In some embodiments, the tip of the implant has a beveled edge. In another embodiment, the tip of the implant has a non-slanted edge.

In some embodiments, after loading the DTMO with the implanting needle, the proximal (posterior) end of the implant is blocked to prevent DTMO from leaking out of the back end of the tube. In another embodiment, the positive pressure / negative pressure adjustment using the plunger of the loading syringe is used to hold the DTMO in the transplantation needle.

As shown in block 7006, a local anesthetic may be optionally administered around the intended implantation site, for example, as is known in the art.

As shown in block 7008, the method may further include positioning the device, including the support structure (Figure 8D), at the implant site such that the support structure abuts the surface of the individual's skin. In some embodiments, the contact between the support structure of the present invention and the surface of the skin of the subject must be airtight so that a vacuum chamber can be formed at a later stage. In one embodiment, the implant is an abdomen of an individual. In other embodiments, the implant is an individual or the like. As another example, the implant may be at another location in the individual's body.

In some cases, the dosage can be adjusted depending on the number / size / efficacy of the DTMO to be implanted. For example, a plurality of DTMOs may be implanted consecutively for a single treatment period to reach a target dosage. In one embodiment, the implant marker template can be used (step 7008) prior to positioning the implant device on the surface of the individual's skin to mark the implant site in multiple places. In one embodiment, the implant marker template is placed over the skin surface of the individual and then displayed on the skin surface to indicate an alignment line for placing the anesthesia line and future support structure 9006, for example. In one embodiment, the surface is marked with a surgical pen or marker. In one embodiment, the surface is marked with a non-permanent dye or ink.

As shown in block 7010, the support structure (Fig. 8D) includes a vacuum chamber with at least one raised protrusion and guide channel 9008, in which a raised protrusion 9007 is adjacent the guide channel . For example, the support structure of FIG. 5A or 5B can be used to maintain and support a skin-related tissue structure for properly implanting DTMO, in order to establish a unique geometric arrangement of the skin, have. For example, applying a vacuum creates a vacuum in the vacuum chamber, causing the skin surface of the skin-related structure to be drawn into the interior of the support structure where the skin-skin layer and dermal layer of the skin- The fat layer can be supported. In an exemplary embodiment, the vacuum state may cause the skin-related structure to be held on the inner support surface of the vacuum chamber. The guide channel may provide guidance and / or stability to the implanted device such that, in one embodiment, proper implantation occurs along a linear implantation axis, such as between the dermal layer and the fat layer, within the subdermal space. In some embodiments, as a result of the implantation, the DTMO remains substantially linear after implantation. In some embodiments, the implantation axis is coaxial with the needle guide channel and the central channel. In another embodiment, the DTMO is implanted without securing linearity.

As shown in block 7012 and Figs. 9A and 9B, by introducing the introducer into the skin-related structure along the implantation axis through the guide channel of the support structure, similar to use in the above-described harvesting method, Can be used to perforate skin-related tissues. This single perforation can be used for any subsequent introduction into the skin-related structure. In this way, damage to the object and scarring are limited. In addition, using the introducer eliminates the problem of exposing the DTMO loaded in the implant to a vacuum while the implant is penetrating the skin-related structure. If the DTMO loaded in the transfer needle is exposed to the vacuum present in the support structure, there is a risk that the DTMO will be inhaled into the vacuum line.

The introducer consisting of the inner needle 9010 and the outer sleeve 9012 fitting as a sheath on the inner needle is inserted into the skin-related structure so that the distal edge of the outer sleeve extends to the tissue region just below the raised protrusion . Under vacuum, the exact geometrical arrangement of the skin-related tissue structure is established such that, upon insertion of the introducer, for example, at a point of penetration, is generally perpendicular to the skin surface. In one embodiment, the inner needle of the introducer and the implant tool have approximately the same diameter dimension. For example, the diameter of the inner needle and the implant may be 10 GA each needle size.

As shown in block 7014 and FIG. 9B, the inner needle is removed from the skin-associated tissue structure, with the outer sleeve member 9012 of the introducer remaining in place. As a result of this operation, the outer sleeve is positioned coaxially with the implantation axis and extends to the skin-associated tissue structure. In a preferred embodiment, the distal edge of the outer sleeve extends up to the fat below the raised protrusion. In another preferred embodiment, the distal edge of the outer sleeve extends to the dermis beneath the raised protrusions.

The outer sleeve may include thin needles, tubes, or any other suitable thin, generally linear object that may be placed in the dermal cavity or subcutaneous space. For example, the outer sleeve may include a needle of 6-18 GA size, e.g., about 10 GA or 14 GA size, as is known in the art. As another example, the outer sleeve may include a plastic tube. In one embodiment, the outer sleeve comprises a high density polyethylene (HDPE) tube. In another embodiment, the outer sleeve comprises a Teflon ^®. In another embodiment, the outer sleeve comprises a polytetrafluoroethylene (PTFE) tube. In another embodiment, the outer sleeve comprises a fluorinated ethylene propylene (FEP) tube.

By inserting at the penetration point at an angle generally perpendicular to the surface of the skin, the introducer can be inserted into the dermis or subcutaneous space. In one embodiment, the inner needle is sloped. In this case, the inner needle part of the introducer can be introduced with the sloped side facing downward. In another embodiment, the sloped side faces up. In another embodiment, the sloped side faces in any direction between upward and downward. In another embodiment, the inner needle is not a slope.

As shown in block 7016 and FIG. 9C, a DTMO 9016-loaded implantation tool 9014 can be inserted through the guide channel and outer sleeve of the support structure, and at a distance approximately equal to the length of the DTMO, Can be advanced along the implantation axis into the layer, the desired site, e.g., the subcutaneous space, the interface of the fat and dermis layers, or the dermal tissue layer. The silicone tube extensions are removed prior to inserting the implant through the needle guide channel, if used to load the DTMO 1716. The transplantation tool can be a needle, such as 6GA-14GA. In one embodiment, the implantation tool can be a 10 GA needle. In other embodiments, the implantation tool may be a 14 GA needle. In one embodiment, the implantation tool is inserted with the slope face up. In another embodiment, the implant tool is inserted with the slope face down. If the implant is advanced to place the DTMO in the desired location, remove the plug if used at the proximal end of the implant.

As shown in block 7018 and in Figure 9D, the stopper tool 9010 and stopper tool 9010 (Figure 8E), including stopper tool body 9018, may be used to secure the implant insert 9016 through the proximal end to the device and the stopper pin To be secured to the device.

As shown in block 7020 and FIG. 9E, the implant needle can be withdrawn, for example, in the subcutaneous space, and the DTMO can be released from the implant needle to place the DTMO linearly along the needle track. In one implementation, to ensure linear placement of the DTMO, the stopper pin 9020 of the stopper tool (Figure 8E) is inserted into the proximal end of the implanting needle. In other embodiments, assistance may be provided to assist in the release of the DTMO, for example, by connecting the syringe to the proximal end of the implant needle and, if possible, evacuating the implant needle, by slowly applying positive pressure to the syringe plunger.

In an exemplary embodiment of the present invention, the stopper of Figure 8E is assembled to a support structure (Figure 8D) such that the load of the stopper is internal and coaxial with the implant. In one implementation, the stopper and support structure are assembled such that the attachment is locked-in-place relative to other members of the implant device, e.g., in that position. For example, the stopper rod may fit within the rear end of the graft needle so that the load is very close to the DTMO loaded in the graft needle, and the graft needle may be withdrawn past the stationary rod of the stopper . The withdrawal of the transplant needle beyond the load, in one embodiment, may be for the full length of the rod. In other embodiments, the withdrawal may be for a portion of the rod length. Because the rod is fixed, with the withdrawal of the needle, the rod may extend past the slope tip of the transplantation needle after withdrawal. In some cases, retracting the graft needle past the stopper rod may prevent the DTMO from being pushed back with the graft needle. In another embodiment, with iron, the DTMO may be released in a linear form into the target site, e.g., subcutaneous space.

Linear transplantation of DTMO may be better for exposing the implanted tissue to the surrounding environment. For example, linear transplantation may be easier to integrate with DTMO. In addition, linear transplantation can facilitate the diffusion of secreted recombinant products, such as recombinant proteins or portions thereof. Furthermore, linear transplantation can promote angiogenesis in the DTMO region. In the future, if a DTMO needs to be removed or removed, a linear arrangement provides a known orientation and location for a given DTMO. In one embodiment, with the implantation, the DTMO is placed linearly in the subcutaneous space. In another embodiment, with transplantation, the DTMO is placed linearly in the same type of tissue as the DTMO, e.g., dermal tissue. In another embodiment, with implantation, the DTMO is placed deeply linearly in the body.

As shown in block 7022, the device can then be removed from the implant. Also, the outer sleeve can be removed at this time.

Those skilled in the art will appreciate that any combination of the above-described operations may be implemented to perform an implant according to embodiments of the present invention. Furthermore, other operations or series of operations may be used.

In addition, in some embodiments of the present invention, without having to repeat all of the embodiments of the apparatus 5000, 6000 described above as a support structure for harvesting a harvesting device, such as a DMO, ) Can also be used for the DTMO transplantation method. That is, in some embodiments of the present invention, device 5000 or 6000 may also be used with implantation methods.

III. Method and apparatus for extracting DMO / DTMO

In accordance with some embodiments of the present invention, systems and methods for in vivo demarcation and localization of implanted dermal micro-organ are provided. Understanding the location of transplantation at a subcutaneous implant site or any other location in the body of a processed tissue, such as a DTMO, is important, for example, if protein therapy should be discontinued or the dose of secreted protein reduced . For example, the administration may be terminated or the dose adjusted by removing one or more of the DTMOs and / or removing one or more of the one, one or more of the transplanted DTMOs. In order to identify the subcutaneously implanted DTMO, an inert, biocompatible ink or dye containing a chromophore, which may be visible to the naked eye or which may require special light emitting conditions to visualize it, Can be colored with DTMO. To this end, the DTMO can be distinguished from surrounding tissue by using visual inspection and / or enhanced imaging means.

In one embodiment, at least the peripheral surface of the DTMO can be coated with, for example, a biocompatible carbon particle, a biocompatible tattoo ink, or other suitable material, including titanium particles, magnetic particles and / or microspheres.

Once implanted subcutaneously, the DTMO can be visualized by the naked eye, by a suitable enhanced imaging device, or by other detection means. Other methods for enhancing the visualization of the implanted DTMO may include using a strong light source on the surface of the skin, or picking up the skin and directing the light source to one side of the skin, whereby the skin appears translucent, DTMO can be seen more clearly. As another example, the dye may be fluorescent, such as a fluorescent plastic bead, visible only when illuminated with UV light.

In other embodiments, the location of the subcutaneously implanted DTMO may be identified by grafting the biocompatible structure together with the DTMO. One example of such a biocompatible structure is a non-absorbable single-stranded nylon suture that is commonly used in numerous surgical procedures. These sutures may be implanted in the same implantation track area as the DTMO, or transplanted so that the spatial location of the DTMO can be confirmed by the suture location, either directly above the DTMO in the upper dermis or directly below the DTMO in the fat. Furthermore, the depth of the DTMO may be known to be at the depth of the subcutaneous space. The suture can be viewed with the naked eye, with the aid of illumination means, and / or with other suitable imaging means such as ultrasound. As another example, the suture may be fluorescent and can be seen in the skin under appropriate UV light. The suture may, in another example, be an absorbent material that can be positioned for a desired period of time, such as months.

In other embodiments, the DTMO can be genetically modified or engineered to contain a fluorescent marker or other marker that can be visualized to include the gene. For example, the DTMO can be modified to have a GFP (green fluorescent protein) gene or a luciferase reporter gene, which can be expressed, for example, with the host of the therapeutic protein. In this manner, the DTMO can be visualized non-invasively using appropriate UV or other suitable illumination and imaging conditions.

In other embodiments, one or more tattoo marks, such as small tattoo points, may be applied to the skin around the implant site. In a preferred embodiment, the small tattoo points are applied to the skin at both ends of the linearly implanted DTMO. Tattoo inks can be permanent or temporary, such as inks used in cosmetic makeup applications.

In some embodiments of the present invention, systems and methods are provided for removing or removing implanted DTMO. For example, if DTMO-based therapy for a patient is to be terminated, or if protein secretion should be reduced, some or all of the transplanted DTMO may be harvested, or some or all of it may be removed. In one embodiment, the DTMO can be surgically removed.

In one embodiment, wherein the skin is applied on both ends of a linearly implanted DTMO with small tattoo points, surgical removal of the DTMO requires at least both tattoo points and the entire skin layer and some subcutaneous tissue , &Lt; / RTI > which can be accomplished by incising an elliptical tissue sample. The incision can then be sutured.

In addition, in some embodiments of the present invention, without repeating both the description of devices 6000 and 1100 (FIGS. 3-6 and 11) described above as a support structure for harvesting a harvesting device, such as DMO, Devices 6000 and 1100 may also be used to extract DTMO.

Hereinafter, FIG. 12 illustrating an exemplary embodiment for extracting DTMO will be described. Those skilled in the art will appreciate that any combination of these harvesting operations may be implemented to perform extraction of DTMO according to embodiments of the present invention. Furthermore, other operations or series of operations may be used.

Hereinafter, all the sampling implementations described in detail above will not be repeated, and will be described with reference to FIG. Briefly, at block 1202, the location of the implanted subcutaneous DTMO can be determined. At block 1203, a local anesthetic may be selectively administered to the site from which DTMO is to be removed. At block 1204, the support structure may be positioned over the location from which the DTMO is to be removed. Using a support structure (e.g., Figure 4C; Figures 5A-B, Figure 6, Figure 11) that can include a vacuum chamber and a guide channel, the skin- So that it can be supported.

At block 1206, a vacuum condition is applied, and skin related tissue structures, including the DTMO to be extracted, can be shaped such that the tissue containing the DTMO to be extracted is in the center channel and coincides with the cutting axis.

At block 1208, a skin-associated tissue can be perforated by inserting the introducer through the guide channel of the support structure into the skin-associated tissue structure at the point of penetration, using an introducer including an inner needle and an outer sleeve. Then, the inner needle is removed, and the outer sleeve is left in a state in which the outer end portion of the outer sleeve is positioned in a region close to the DTMO to be extracted.

At block 1210, tissue cores containing DTMO may be recovered. A coring needle (eg, 11 GA or similar) with a diameter equal to or greater than the DMO sampling needle can be inserted through the guide channel and outer sleeve along the cutting axis to retrieve the already implanted DTMO. In one embodiment, the DTMO is removed using a coring tube of diameter equal to or greater than that used to directly pick DMO. In one embodiment, additional tissue surrounding the DTMO being removed is retrieved during extraction of the DTMO. In one embodiment, the additional tissue comprises epidermal tissue. In one embodiment, the additional tissue comprises dermal tissue that is not combined with DTMO. In one embodiment, the additional tissue comprises adipose tissue. In one embodiment, this coring approach can be combined with vacuum suction at the proximal end of the cutting tool to aid in retrieving tissue samples incised from the body.

In an embodiment of the invention, a method of minimally invasive or non-invasive removal of DTMO in an in-drilling can be used, with little or no trauma to the patient, and little invasiveness. In one embodiment, a laser, such as a non-invasive Yag laser, can potentially be used in conjunction with the case of a dyed DTMO. The energy of the Yag laser can be selectively absorbed by the chromophore of the dyed DTMO, for example, so that energy is mainly directed to the DTMO, resulting in minimal damage to the surrounding tissue. Other light energy sources may be used. As another example, this optical scheme can be used with other means of identifying the DTMO position other than using a dye.

In another embodiment, the DTMO may be removed by transferring breakdown energy from a minimally invasive probe inserted into the subcutaneous space along the length of the DTMO. These probes can carry various energy types such as radio frequency, cryogenic temperature, microwave, and heat. The implanted structures, such as sutures, can be used to identify the location of the DTMO, which allows the probe to be inserted subcutaneously, for example, just above or just below the suture, or along the suture. In this case, for example, the breaking energy can be delivered with the suture still in place. As another example, the suture can be removed after placing the probe and before delivering the fracture energy. The amount of energy may be necessary to denature proteins in tissues, such as during coagulation by diathermy. Additionally or as an alternative, the amount of energy applied may be as much as that used in surgical cutting electronics that burn tissue. Of course, other means of locating the fracture energy and other means of delivering the fracture energy may be used.

IV. Method and apparatus for processing DMO

The DMO is selectively genetically modified, e. G., In accordance with embodiments of the present invention. Methods and apparatus for processing DMO are described in detail in U.S. Patent Application Publication No. US-2012/0201793-A1, the entire contents of which are incorporated herein by reference.

In one embodiment, the invention provides a method of delivering a gene product of interest to a subject by transplanting the genetically modified DMO of the invention to the subject.

In one aspect, the invention includes the use of genetically modified DTMOs for transplanting into organisms. The terms "administering "," transplanting ", and "transplanting ", as used herein, may be used interchangeably and refer to the localization of DTMO at the desired site , It may refer to placing the DTMO of the present invention in an individual, e.g., a self, a homogeneous or heterogeneous individual. The DTMO is implanted at a desired location on the individual in such a way that at least some of the cells of the DTMO remain viable. In one embodiment of the invention, at least about 5%, at least about 10% in another embodiment of the invention, at least about 20% in another embodiment of the invention, in another embodiment of the invention, at least about 30% %, In another embodiment of the invention, at least about 40%, and in another embodiment of the invention, at least about 50% or more of the cells remain viable after administration to the subject. The survival period of cells after administration to an individual may be several hours, such as a few hours, such as 24 hours to several days, as long as several weeks to months or even years.

In another example, a DTMO, including genetically modified cells, can be maintained in vitro and the therapeutic agent remaining in the media supernatant surrounding the tissue sample can be isolated and injected or applied to the same or different entities.

Alternatively or additionally, the DTMO may be prepared by a method known in the art immediately after or immediately after genetically modified from a tissue sample, such as, but not limited to, a gradual increase in DEME containing 10% DMSO (0 deg. C, -20 deg. C, -80 deg. C, -196 deg. C) and cryogenic storage can be performed.

According to one aspect of some embodiments of the present invention, the number of DTMOs to be implanted is determined by one or more of the following: daily administration to a subject based on normal guidelines, specific clinical protocols, or demographics of similar entities The corresponding amount of the subject therapeutic agent. The corresponding amount of therapeutic agent, such as the subject protein, specific for the same individual, if he or she has been previously administered by infusion or other route. Object data such as weight, age, physical condition, and clinical condition. The pharmacokinetic data of previous tissue samples, including the administration of genetically modified cells to other similar individuals. Individual response to previous DTMO administration.

In terms of some embodiments of the present invention, only some of the DTMOs are used in a given treatment period. The remaining DTMOs may be forwarded and maintained (or cryogenic or otherwise stored) for future use.

Also in accordance with an embodiment of the present invention there is provided a method of treating a disorder in a subject, comprising: (a) monitoring the level of the therapeutic agent in the subject; (b) comparing the level of the agent to the desired level; (c) if the level is below a minimum level, further implanting the DTMO; And (d) removing or extracting one or more transplanted DTMOs if the level is higher than the highest level, adjusting the dose of the therapeutic agent produced by the DTMO implanted in the subject, and releasing the therapeutic agent &Lt; / RTI > Optionally, the method comprises a periodic repetition of (a) - (d). As another example or additionally, removal or extraction consists of removing or removing a portion of the implanted one or more DTMOs. Optionally, the extraction includes surgical excision. Optionally, removal includes killing a portion of the implanted DTMO.

As described above with reference to FIG. 1, at least some of the processes of maintaining DMO at the time of genetic modification, as well as genetic modification itself, can be performed in the bioreactor.

Example

Example 1

Collection of dermis micro-organ

Dermal micro-organ was collected from human subjects under sterile conditions.

Experimental course

In consideration of the easiness of the individual, the lower abdomen was selected as the harvesting section, sterilized and marked with a guide line, and local anesthetics were injected. The harvesting area was a healthy skin area without scratches or other obvious skin abnormalities. 4C) structure with a vacuum control hole was connected to a vacuum source and the vacuum was activated, and then the support structure was placed in the selected collection of the entity skin without covering the vacuum control hole.

The vacuum control holes were closed with fingers to create a vacuum so that the skin-related tissue structure rises to the vacuum chamber.

The introducer 4006 and 4008 were quickly inserted into the needle guide of the support structure until the sharp slope point of the needle inside the introducer was directed downward (Fig. 4D, 4006) until it was completely stopped. Then, the inner needle 4006 of the introducer was taken out and the introducer sleeve 4008 was left.

Next, the sharp tip of the coring needle attached to the medical drill was slowly inserted into the introducer sleeve in a forward direction until the tip reached the distal end of the sleeve. Then the drill was actuated and pushed forward until it was completely stopped, and the tip of the coring needle was pushed through the dermal tissue into the adipose tissue. At this time, the finger was removed from the vacuum control hole to inactivate the vacuum.

Thereafter, the drill is detached from the coring needle, the collet is slid onto the needle and is then pushed over the needle, and the septum is pierced through the exposed end of the coring needle and is then connected to the collector so that the needleless valve syringe assembly, Lt; / RTI > The syringe was connected to the bulkhead and the syringe was retracted with the coring needle while pulling the plunger of the syringe to form a vacuum. During this pulling process, the DMO was aspirated into the syringe body (Figure 3G).

Experiment result

Several DMOs were collected. The extracted DMO is compared with a toothpick in Figure 14A, where the DMO is about 30 mm long. As shown in Fig. 14B (1402), scar occurrence was minimized in the skin tissue of the harvesting portion.

Example 2

Transplantation of dermal micro-organ

The dermal micro-organ was implanted into human subjects under sterile conditions.

Experimental course

Similar to the preparations for harvesting, the transplantation of the dermis micro-organ into the human subject began with ease to the individual, the lower abdomen was selected as the graft, the guide for sterilizing and then aligning the support structure for transplantation, Local anesthetics were injected at that location. A vacuum source was connected to a sterile implant support structure with a vacuum control hole (Fig. 8D), the vacuum power was turned on, and the support structure was placed on the transplantation site marked on the epidermis of the individual without covering the vacuum control hole .

A vacuum control hole was covered with a finger to induce a vacuum so that the skin-related tissue structure was sucked into the vacuum chamber.

The introducer was quickly inserted into the needle guide of the support structure, with the sharp slope point of the needle inside the introducer pointing down (Fig. 8C), until it was completely stopped. Then, the inner needle of the introducer was taken out and the introducer sleeve was left.

Next, the transfer needle into which the DMO was injected at the distal end was inserted into the introducer sleeve, and was pushed in the forward direction until it was completely stopped. The stopper member was then connected to the implant by inserting a stopper pin into the inner lumen of the implant. The stopper pin was moved in the forward direction to be positioned very close to the DMO injected into the implant needle. The stopper body was secured to the implant structure and retracted past the stopper pin and the stopper pin was held in place while the DMO was linearly implanted in the subcutaneous space of the implant.

The implanted device was carefully removed from the implant site, and the finger was removed from the vacuum hole to release the vacuum. To indicate the location of the implantation site, both ends of the linearly implanted DMO on the skin surface were marked with tattoo points with semi-permanent inks.

Experiment result

Several DMOs were transplanted. As shown in FIG. 14B, a DMO was implanted in the lower abdomen 1404, and the transplantation site was confirmed as a tattoo point. At the graft site, scarring of skin tissue was minimized.

While the invention will be described by way of illustrative and non-limiting embodiments with reference to the embodiments, it will be obvious that it is not intended to limit the scope of the invention. For example, a limited number of gene mutations are described. However, it is believed that all of the substantial genetic changes in the tissue, induced by any of the known methods, based on the methods described herein for re-implanting live tissue into the body of a patient, and the body's survival of the tissue after insertion, It is evident that the patient will cause secretion of the target protein or other therapeutic agent.

Various implementations of the present invention may be made by those skilled in the art, including combinations of features of various implementations. The scope of the present invention is limited only by the claims. Also, to avoid any doubt as to the scope of the claims, when " comprising ", "comprising" or "having" and their transitional terms are used in the claims, Quot ;, " includes ".

Also, in this application, the term "comprising " includes elements in which the system is mentioned, but is not meant to exclude other elements that may be optional. The expression " consisting essentially of "means a method that includes the referenced elements but excludes other elements that may have an inherently significant effect in performing the method. In other words, "consisting of" would mean excluding more elements than traces. Implementations defined by each of these variation terms are included within the scope of the present invention.

Further, in the present application, the term " about "means a variation of 0.0001-5% in the numerical range or numerical range mentioned. In one embodiment, the term " about "means the number or variance of 1 - 10% in the numerical range mentioned. In one embodiment, the term " about "means a deviation of up to 25% in the number or range of values mentioned.

Further, in the present application, the term "a, one or an " refers to one or more. In one embodiment, the expression" two or more "may be any denomination for a particular purpose

In addition, in the present application, the term "treatment" refers to both therapeutic treatment and prophylactic or preventative measures, the purpose of which is to prevent or attenuate the desired pathological symptoms or disorders. Thus, in one embodiment, the treatment may include a direct effect on the disease, disorder or condition or cure, inhibition, inhibition, prevention, severity relief, onset delay, symptom relief, or combinations thereof. Thus, in one embodiment, "treating" refers in particular to progressive delay, rapid remission, induction of remission, remission enhancement, acceleration of recovery rate, increased efficacy or tolerance of alternative therapies, or combinations thereof. In one embodiment, "preventing" refers in particular to delaying the onset of symptoms, preventing the recurrence of the disease, reducing the frequency or frequency of recurrences, prolonging the latency between onset of symptoms, or a combination thereof. In one embodiment, "inhibiting" or "inhibiting" is especially effective in reducing the severity of symptoms, decreasing the severity of acute events, decreasing the number of symptoms, decreasing the incidence of disease-related symptoms, Secondary infection reduction, prolonged patient survival, or a combination thereof.

While certain features of the invention have been illustrated and described herein, many modifications, substitutions, changes, and equivalents will now occur to those skilled in the art. It is to be understood that the appended claims are intended to cover all such modifications and changes as fall within the true spirit of the invention.

Claims

An apparatus for harvesting a dermal micro-organ,
The device
a. A support structure for supporting a skin-related tissue structure from which the dermal micro-organ is to be harvested;
b. Introducer; And
c. A cutting tool,
Wherein the support structure comprises:
(a) a first tubular element comprising an insert for the device;
(b) a vacuum chamber comprising an inner support surface and two raised protrusions, which are elevated protrusions relatively distant from the insert and distant protrusions; And
(c) a central channel positioned between the two raised protrusions,
The raised protrusions can support and fix the skin-related tissue structure in a preferred form and position so that the cutting tool can cut the dermal micro-organ from the skin-related tissue structure under vacuum,
The first tubular member being connected to the vacuum chamber and one or more vacuum channels to fluidically couple the vacuum chamber with one or more vacuum sources;
Wherein the central channel supports the skin related tissue structure such that when the cutting tool is inserted into the first tubular member, the dermal layer of skin is located within the trajectory of the cutting tool,
Wherein the dermis micro-organ is essentially composed of a plurality of dermis components, and wherein the complete epidermal layer is absent.

The method according to claim 1,
Wherein the first tubular member is adapted to coaxially align the cutting tool into the central channel.

The method according to claim 1,
Wherein the introducer comprises a second tubular member and a fourth tubular member,
a. The second tubular member being inserted into the fourth tubular member and extending beyond the distal end of the fourth tubular member;
b. Said second tubular member and said fourth tubular member being coaxially inserted together into said first tubular member at said insert;
c. Wherein the fourth tubular member is coaxial with the first tubular member upon withdrawal of the second tubular member.

The method of claim 3,
Said second tubular member comprising a needle,
And said fourth tubular member comprises an outer sleeve.

The method of claim 3,
Wherein the cutting tool is inserted into the fourth tubular member.

The method according to claim 1,
Wherein the cutting tool comprises a third tubular member insertable in the insert through the fourth tubular member coaxially remaining in the first tubular member,
Wherein the cutting tool is substantially coaxial with the first tubular member,
Wherein the third tubular member is capable of cutting the dermal micro-organ from the skin-associated tissue structure.

The method according to claim 6,
Wherein the third tubular member comprises a coring tube capable of cutting through the skin related tissue structure when advanced along a cutting axis,
Wherein the cutting axis is substantially coaxial with the first tubular member.

8. The method of claim 7,
Wherein the core ring tube comprises a rotatable core ring tube attached to a power source.

9. The method of claim 8,
Wherein the power source is selected from an electric motor or an air-driven turbine.

The method according to claim 1,
Wherein the core ring tube is a coring needle having a symmetrically sharpened tip.

A method for collecting dermal micro-organ from an individual,
The method comprises:
a. Placing the device according to claim 1 in contact with the surface of the skin of the subject and placing it in the collection section;
b. Supporting a skin-related tissue structure from which the dermal micro-organ is to be harvested;
c. Drilling the skin related tissue structure;
d. Cutting the dermal micro-organ from the supported skin-related tissue structure; And
e. And recovering the dermal micro-organ,
Wherein the dermal micro-organ is essentially constituted by a plurality of dermal components and wherein the complete skin layer is absent.

12. The method of claim 11,
Wherein the sampling method comprises making only one puncturing point in the skin-related structure.

12. The method of claim 11,
The apparatus comprises:
a. A support structure for securing the skin-related tissue structure from which the dermal micro-organ is to be harvested;
b. A introducer comprising a second tubular member and a fourth tubular member; And
c. And a cutting tool including a third tubular member insertable into the guide channel and the outer sleeve,
Wherein the support structure comprises a first tubular member and a vacuum chamber, wherein the first tubular member is a guide channel substantially parallel to the skin surface of the entity, the guide channel is connected to the vacuum chamber, Includes an inner support surface and two raised protrusions,
Wherein the second tubular member comprises an inner needle and the fourth tubular member comprises an outer sleeve, the inner needle being inserted into the outer sleeve and extending beyond the distal end of the outer sleeve,
Wherein the third tubular member comprises a cutting tool that is a coring tube for cutting the dermal micro-organ from the skin-associated tissue structure.

12. The method of claim 11,
a. Wherein the supporting step comprises applying a vacuum to the vacuum chamber, wherein the vacuum condition causes the skin-related structure to be secured to the inner support surface of the vacuum chamber;
b. The perforating may include inserting the introducer through the guide channel into the skin-related structure, and then withdrawing the inner needle from the outer sleeve and the skin-related structure, wherein the outer sleeve comprises the guide channel and the skin- &Lt; / RTI >
c. Wherein cutting comprises coaxially inserting the coring tube into the outer sleeve and inserting the coring tube into the skin related structure.

15. The method of claim 14,
Wherein the recovering comprises using a vacuum to recover the dermal micro-organ into the enclosed container in the coring tube.

15. The method of claim 14,
Wherein said recovering step comprises pushing said dermal micro-organ through said coring tube.

15. The method of claim 14,
Wherein the cutting step comprises:
a. Rotating the coring tube such that the coring tube advances toward the distal end of the apparatus; And
b. Further comprising withdrawing the coring tube from the skin-related structure while applying a vacuum to the coring tube, thereby withdrawing the dermal micro-organ into the enclosed container.

As a device for transplanting a dermal micro-organ,
The device
a. A loading syringe comprising a first tubular member;
b. An implantation tool comprising a second tubular member;
c. A support structure for locating the skin related tissue structure to which the dermal micro-organ will be implanted;
d. A introducer for perforating the skin-related tissue structure;
e. And a stopper tool connectable to the support structure,
The stopper tool includes a rod, which assists in maintaining the position of the dermal micro-organ during implantation;
Wherein the support structure comprises a third tubular member comprising an insert of the support structure,
Said second tubular member being insertable into said third associated member,
Said third tubular member providing an implantation axis,
Wherein the dermal micro-organ is essentially constituted by a plurality of dermal components and wherein the complete skin layer is absent.

19. The method of claim 18,
And the third tubular member is a guide channel.

21. The method of claim 20,
a. A vacuum chamber including an interior support surface capable of securing the skin-associated tissue structure in a preferred form and position so that the implantable device can implant the dermal micro-organ into the skin-associated tissue structure; And
b. And one or more vacuum channels for fluidly coupling the vacuum chamber with one or more vacuum sources,
And the third tubular member is connected to the vacuum chamber.

21. The method of claim 20, wherein the vacuum chamber
a. One or more raised protrusions capable of supporting a plateau comprising a skin epidermal layer and a skin dermis layer derived from said skin-associated tissue structure on the orbit of said implant; And
b. Further comprising a central channel coaxial with said third tubular member and remote from said elevated protrusion relative to said insert,
Wherein the central channel is capable of supporting skin-related tissue.

20. The method of claim 19, wherein the introducer
a. A fourth tubular member, and
b. A fifth tubular member,
The fourth tubular member being inserted through the fifth tubular member and extending beyond the proximal end of the fifth tubular member;
Said fourth tubular member and said fifth tubular member being coaxially inserted into said third tubular member at said insert,
Wherein the fifth tubular member remains in the third tubular member coaxially with the third tubular member upon withdrawal of the fourth tubular member.

23. The method of claim 22,
The fourth tubular member is an inner needle,
And said fifth tubular member is an outer sleeve.

20. The method of claim 19,
Wherein the second tubular member comprises a graft needle that is advanced along the graft axis and is capable of grafting dermal micro-organ along the graft axis.

19. The method of claim 18,
Wherein the implantation tool comprises a beveled tip.

19. The method of claim 18,
Wherein the dermal micro-organ is a genetically modified dermal micro-organ.

As a method for transplanting dermal micro-organ into an individual,
The method comprises:
a. Loading the dermal micro-organ into a loading syringe comprising a first tubular member;
b. Moving the dermal micro-organ from the loading syringe to an implantation tool comprising a second tubular member;
c. 20. A method comprising: disposing an apparatus according to claim 19 at a transplantation site, wherein the apparatus is in contact with a skin layer of the subject, the transplantation axis being at a right angle to the skin epidermal layer of the subject;
d. Supporting at the implantation site a skin-related tissue structure to which the dermal micro-organ is to be implanted;
e. Drilling the skin related tissue structure;
f. Advancing the implant tool into the skin-associated tissue structure along the implantation axis;
g. Withdrawing said second tubular member and retaining said dermal micro-organ in said skin-associated tissue structure,
Characterized in that the dermal micro-organ consists essentially of a plurality of dermal components and the complete skin layer is absent.

28. The method of claim 27,
Wherein the dermal micro-organ is a genetically modified dermal micro-organ.

28. Apparatus according to claim 27,
a. A loading syringe comprising a first tubular member;
b. An implantation tool comprising a second tubular member;
c. A support configuration for supporting the skin related tissue structure to which the dermal micro organ is to be implanted;
d. A introducer comprising a fourth tubular member and a fifth tubular member; And
e. And a stopper tool including a load,
Wherein the support structure comprises a third tubular member and a vacuum chamber, the third tubular member being substantially parallel to the skin surface of the entity, the third tubular member and the vacuum chamber defining a second tubular member A transplantation tool including the first tubular member and the second tubular member is coaxially implanted into the third tubular member;
The fourth tubular member being inserted into the fifth tubular member and extending beyond the distal end of the fifth tubular member;
When the stopper tool is engaged with the device, when the rod is inserted into the third tubular member, the rod is coaxially aligned with the second, third and fifth tubular members,
Wherein at least a portion of the rod is inserted into the second tubular member.

30. The method of claim 29,
Wherein the vacuum chamber comprises at least one raised protrusion and a central channel,
Wherein the raised protrusion is proximate to the insertion site, the center channel is remote from the insertion,
Wherein the vacuum condition aligns the implantation axis coaxially with the center channel.

28. The method of claim 27,
a. Wherein the supporting step comprises applying a vacuum to the vacuum chamber, wherein the vacuum condition causes the skin-related structure to be fixed at the inner support surface of the vacuum chamber,
b. Wherein said perforating comprises inserting said fourth and fifth tubular members substantially coaxially through said third tubular member into said skin related structure and then removing said fourth tubular member from said skin- Withdrawing the member and leaving the fifth tubular member in the third tubular member and the skin related structure,
c. Wherein the implanting includes inserting the second tubular member coaxially through the fifth tubular member and inserting the second tubular member into the skin related structure;
d. The withdrawing step withdrawing the second tubular member on the rod, wherein the rod member is held stationary relative to the support structure, and wherein the micro-organ is implanted into the entity through the withdrawal How to.

32. The method of claim 31,
Wherein only one puncturing point is formed in the skin-related structure at the time of insertion.

32. The method of claim 31,
Wherein the implanting step comprises implanting the dermal micro-organ into a liner form.

32. The method of claim 31,
RTI ID = 0.0 > 1, < / RTI > wherein said transplantation is implantation into dermal tissue.

32. The method of claim 31,
Wherein said implantation is implantation into the skin or under the skin.

32. The method of claim 31,
RTI ID = 0.0 > transplantation. &Lt; / RTI >