WO2019013533A2 - Method for manufacturing blended filament yarn for cell culture support, blended filament yarn for cell culture support realized therethrough, and fabric comprising same - Google Patents

Abstract

Provided is a method for manufacturing a blended filament yarn for a cell culture support. The method for manufacturing a blended filament yarn according to one embodiment of the present invention is performed by comprising the steps of: (1) preparing a first yarn having at least one of a first motif and a first motif conjugate having an amino acid sequence in which a first amino acid sequence motif is repeated at least twice, and a second yarn having at least one of a second motif, which is different from the first motif, and a second motif conjugate having an amino acid sequence in which a second amino acid sequence motif is repeated at least twice, respectively; and (2) plying the prepared first yarn and second yarn. According to this, different motifs are provided in a unitary blended filament yarn, and the blended filament yarn is very suitable for fully expressing each function of the motifs. Also, such a unitary blended filament yarn may make it possible to implement a micro environment in which the steps of attachment, migration, proliferation, and differentiation of cultured cells can take place continuously. Furthermore, it is possible to culture cell aggregates formed of cells with the same differentiation step, and at the same time to culture multiple cell aggregates with different differentiation steps through a unitary blended filament yarn. In addition, it is possible to three-dimensionally grow the cultured cells to be more suitable for application for in vitro experimental models or implantation in animal bodies, and thus the present invention may be widely applied to various products used in the field of cell culturing or tissue engineering such as a bioreactor, a cell culture container, an implantation kit, and the like.

Description

Method for manufacturing horn sheath for cell culture support, horn sheath for cell culture support embodied thereby and fabric

The present invention relates to a heterogeneous cell for cell culture support, and more particularly, to a micro-environment in which the process of attachment, migration, proliferation, and differentiation of cells cultured through a single hornblende can be continuously performed, A method for producing the same, and a fabric comprising the same.

Recently, as the use of cultured cells for disease treatment is expanded, interest and research on cell culture are increasing. Cell culture is a technique for extracting cells from a living body and culturing them out of the living body. The cultured cells are differentiated into various tissues of the body such as skin, organs, nerves, etc. and transplanted into the human body before transplantation into the human body. And can be used for treating various diseases.

This field of tissue culture is related to cell culture and it is a multidisciplinary study applying existing scientific fields such as cytology, life sciences, engineering, medicine, etc., and the correlation between the structure and function of living tissue And new fusion techniques are being studied to replace damaged tissues or organs with normal tissues and regenerate them.

One of the tasks that have received much interest and research / developments in the field of conventional cell culture or tissue engineering using the same is to use a material of a support which can be cultured / differentiated and implanted into a human tissue with cells, Structure, and so on.

In recent years, techniques for culturing cells by introducing substances beneficial for cell culture in a cell culture culture environment have been introduced. For example, the method of providing the beneficial substance in a culture medium is advantageous in that the material advantageous to the cell culture is difficult to be continuously disposed near the cells attached to the support as the substance flows in the culture solution, There is a difficult problem.

In order to compensate for this, a method of culturing a cell by providing a material advantageous for cell culture to a cell culture supporter has been disclosed. This method can be performed at any stage of culturing the cell, for example, cell adhesion / proliferation and differentiation There is a problem that it is difficult to promote the two steps at an appropriate time. In order to solve this problem, when the adhesion / proliferation promoting substance and the differentiation promoting substance are both provided on one support, the differentiation of the proliferated cells occurs simultaneously during the proliferation of some cells in the proliferated one group, There is a problem that different cells are included in one group of cells. In addition, when a substance advantageous to two or more kinds of cell cultures is included, the chemical and / or physical bonding between the two substances occurs in the process of providing them to the cell culture support, so that their respective original functions are not completely expressed, There is a problem that can not be done.

Therefore, even if a support is provided with two or more kinds of cell culture beneficial substances, the respective functions are fully expressed, and the differentiation steps are performed on the same cells It is an urgent need to study a support for cell culture which can culture cell aggregates formed at a plurality of different cell aggregation stages simultaneously and can further promote cell culture.

SUMMARY OF THE INVENTION The present invention has been made in view of the above-mentioned problems, and it is an object of the present invention to provide a method and apparatus for preventing the chemical and / And to provide a method for producing the horn sheath for cell culture support and a method for producing the same.

The present invention also provides a method for preparing a hornblende for a cell culture support and a method for producing the same which can realize a microenvironment in which the processes of attachment, migration, proliferation and differentiation of cells cultured through a single hornblende can be continuously performed There is a purpose.

The present invention also relates to a method for producing a hornblende for cell culture support, which is capable of culturing cell aggregates formed of cells having the same differentiation stage through a single horny filament and culturing a plurality of cell aggregates having different differentiation stages, There are other purposes to provide.

It is another object of the present invention to provide an in vitro experimental model or a hybrid seed support for a cell culture support capable of three-dimensionally growing cells cultured so as to be more suitable for implantation in an animal body.

In addition, the present invention can stably cultivate the cells even in a culture condition or a vibrating condition in which the cell culture liquid flows, and the sufficient mechanical strength is developed so that collapse of the support caused in the cell culture process can be prevented, Another object of the present invention is to provide a horny sheath for a cell culture support and a fabric for a cell culture support comprising the horny sheath for a cell culture support which can be stably grown as a three dimensional cell cluster.

In addition, the present invention provides a fabric for a cell culture support which can be widely applied to various products used in the field of cell culture or tissue engineering such as a bioreactor, a cell culture container, There is another purpose.

In order to solve the above-mentioned problems, a first embodiment of the present invention is a method for producing a first motif, comprising: (1) preparing a first motif and a first motif having at least one of a first motif conjugate having an amino acid sequence in which a first motif amino acid sequence is repeated at least twice, Preparing a second yarn having at least one of a yarn and a second motif conjugate having an amino acid sequence in which the second motif is different from the first motif and the second motif amino acid sequence is repeated at least twice; 2) folding the prepared first yarn and the second yarn into a cell culture support.

According to an embodiment of the present invention, the first yarn and the second yarn may each independently include a mono yarn, which is a yarn, a filament yarn, or a slitting yarn, as one strand or a plurality of strands.

The monosaccharide may be selected from the group consisting of polystyrene (PS), polyester, polyether sulfone (PES), polyvinylidene fluoride (PVDF), polyacrylonitrile (PAN), polydimethylsiloxane (PDMS) Poly (alkylene oxide), poly (amino acids), poly (allylamines), polyphosphazene, and polyethylene oxide-polypropylene oxide block copolymers. Polyglycolic acid, PLLA (poly (L-lactide)), PLGA (poly (lactic acid)), polyglycolic acid, polyglycolic acid, DL-lactide-co-glycolide), polylactic acid, and polyvinyl alcohol as a fiber-forming component.

In addition, the first yarn and the second yarn may be partially twisted after twisting or twisting.

In addition, the first yarn or the second yarn of the mixed yarn may not include at least one first motif and at least one second motif in a chemically or physically bonded motif combination.

The first yarn or the second yarn may independently have a fineness of 0.01 to 300 denier.

The slitting yarn may be a fibrous web of a three-dimensional network structure cut to have a predetermined width. At this time, the fibrous web may have a basis weight of 0.1 to 100 g / m 2 and a width of 0.1 to 30 mm.

Further, the fibrous web may have a ratio of basis weight to thickness of 1: 0.8 to 1.7.

The fibrous web may be a nanofiber web having a basis weight of 7 to 30 g / m 2 and a thickness of 7 to 30 탆.

In addition, the first motif and the second motif may have a function of promoting at least one of attachment, migration, proliferation and differentiation of cells.

According to a first aspect of the present invention, there is provided a method for producing a microorganism having a first motif and a first motif having at least one of a first motif and a first motif conjugate having an amino acid sequence in which a first motif amino acid sequence is repeated at least twice, The present invention provides a hybrid filament for cell culture support comprising a first filament and a second filament having at least one of a first motif, a heterologous second motif, and a second motif conjugate having an amino acid sequence in which a second motif amino acid sequence is repeated at least twice .

According to an embodiment of the present invention, the first yarn or the second yarn of the mixed yarn may not include a motif combination body in which at least one first motif and at least one second motif are chemically or physically combined.

In order to solve the above-mentioned problems, a second embodiment of the present invention is a method for producing a yarn comprising a first yarn having a supporting function, and a second yarn covering at least a part of the outer surface of the first yarn, The present invention provides a hornblende for cell culture support comprising a second yarn.

According to an embodiment of the present invention, the first yarn is a multi yarn having a single yarn or a plurality of monosa, and may have an average diameter of 100 to 1000 mu m.

In addition, when the first yarn is multisignal, the number of strands may be 2 to 10.

The mixed yarn may be formed by covering the outer surface of the first yarn with a second yarn having a slitting yarn formed by cutting a nanofiber web having a three-dimensional network structure formed of nanofibers into a predetermined width. At this time, the slitting yarn may have a basis weight of 5 to 30 g / m 2, a thickness of 7 to 30 탆, and a ratio of the basis weight to the thickness of 1: 0.8 to 1.7.

In addition, the first yarn may be a multi-yarn having a plurality of mono yarns in order to have a three-dimensional surface morphology.

The first yarn and the second yarn may be present in a weight ratio of 1: 0.5-10.

The second yarn may be a nanofiber aggregate formed of nanofibers electrospun on the outer surface of the first yarn.

The second yarn may also be nanofibers electrospun on the outer surface of the first yarn.

In addition, the nanofiber aggregate may include a surface portion on which the nanofibers cover the outer surface of the first yarn, and a protrusion formed on the surface portion and protruding from the nanofibers toward the outside.

The first yarn may be a multi-yarn having a plurality of mono yarns in order to improve the adhesion between the nanofiber aggregate and the first yarn.

The thickness of the surface portion may be 0.2 to 5 탆.

In order to improve the adhesion between the surface portion and the first yarn, some of the nanofibers of the surface portion may be partially or wholly bonded to each other to cover the outside of the first yarn.

The weight ratio of the first yarn and the nanofiber aggregate may be 1: 2 to 5.

The first yarn or the second yarn may independently have a fiber forming component selected from the group consisting of polystyrene (PS), polyester, polyether sulfone (PES), polyvinylidene fluoride (PVDF), polyacrylonitrile , Polydimethylsiloxane (PDMS), polyamide, polyalkylene, polyalkylene oxide, polyurethane, poly (amino acids), poly (allylamines), poly At least one non-biodegradable component selected from the group consisting of polyphosphazene and polyethylene oxide-polypropylene oxide block copolymer, or at least one of a polycaprolactone, a polydioxanone, a polyglycolic acid, , Polylactic acid, and polyvinyl alcohol), polylactic acid (PLA), poly (L-lactide), PLGA Ingredients .

In addition, the second source may have a physiologically active component that induces at least one of cell adhesion, migration, growth, proliferation, and differentiation.

In addition, the physiologically active component may be selected from the group consisting of monoamines, amino acids, peptides, saccharides, lipids, proteins, glucoproteins, glucolipids, proteoglycans, mucopolysaccharides, ) And one or more of the cells.

The present invention also provides a fabric for a cell culture support comprising a hornblende yarn according to the present invention.

The present invention also provides a tissue engineering implant comprising a fabric according to the present invention and cells cultured in the fabric.

At this time, the cell may be any one or more stem cells selected from the group consisting of ovarian stem cells, pluripotent stem cells, pluripotent stem cells, oligopotent stem cells, and single stem cells, and hematopoietic stem cells, hepatocytes, The present invention can include at least one of differentiated cells selected from the group consisting of cells, mesothelial cells, endothelial cells, muscle cells, nerve cells, immune cells, adipocytes, cartilage cells, bone cells, blood cells and skin cells.

Hereinafter, terms used in the present invention will be described.

The term " extracellular matrix (ECM) " of the present invention is a substrate surrounding the outside of a cell, which means that it occupies cells and cells and has a network structure mainly composed of proteins and polysaccharides.

The " motif " of the present invention can be structurally / functionally interacted with receptors contained in proteins or glycoproteins in the extracellular matrix, which play an important role in cell adhesion, migration and differentiation, Or a peptide comprising an amino acid sequence as shown in SEQ ID NO: 1, which is isolated in a cell or artificially produced using a gene cloning technique.

The term " 3dimension cell cluster " of the present invention means a three-dimensional structure in which cells are three-dimensionally gathered.

In the " heterologous " motif of the present invention, heterologous means that the amino acid sequences are different from each other and / or the functions are different from each other.

According to the present invention, the production method according to one embodiment of the present invention is well suited for providing a single motif in a single hybrid filament yarn, and the motif function of each of them is fully expressed. In addition, a single hybrid fiber can realize a micro environment in which the process of attachment, migration, proliferation, and differentiation of cultured cells can be continuously performed. Furthermore, it is possible to cultivate cell aggregates formed with the same cells in differentiation stages through a single hornblende, and to cultivate various cell aggregates having different stages of differentiation.

In addition, the hybrid fiber for a cell culture support can improve the survival rate and proliferation rate of cells and form a support capable of sterically multiplying cells, or can function as a support by itself. Furthermore, mechanical properties such that the cells can stably multiply in a three-dimensional cell cluster can be ensured even in a culture condition or a vibration condition in which the cell culture fluid flows.

In addition, it is possible to sterically grow cells cultured to be more suitable for in vitro experimental models or transplantation in an animal body, and can be used for cell culture or tissue engineering fields such as bioreactors, cell culture containers, And can be widely applied to various products used in the art.

FIGS. 1 to 4 are a perspective view and a partially enlarged view illustrating various embodiments of a horn sheath according to an embodiment of the present invention;

5 is a perspective view and a partially enlarged view of a mixed fiber according to an embodiment of the present invention,

FIG. 6 is a perspective view of a mixed fiber for a cell culture support according to an embodiment of the present invention,

FIGS. 7A and 7B are various embodiments of first yarns constituting the horny sheath according to an embodiment of the present invention. FIG. 7A is a perspective view of a first yarn, which is a multifilament multifilament yarn, 7B is a perspective view showing that a part of the twisted first yarn of FIG. 7A is partly broken to include a space between the strands constituting the first yarn,

8 is a cross-sectional view of a mixed fiber for a cell culture support according to an embodiment of the present invention,

9A to 9C are SEM photographs of a mixed fiber for a cell culture support in which a second yarn is directly electrospun on a plurality of strands of a first yarn according to an embodiment of the present invention, 9C is a photograph enlarged to x 5.0 k times,

10A and 10B show an example of a slitting yarn included in an embodiment of the present invention. FIG. 10A is an enlarged photograph of a state of a fiber web before it is made into a slitting yarn, FIG. 10B is an enlarged photograph after being made of a slitting yarn,

11 is a photograph showing an intermediate step for manufacturing a slitting yarn included in an embodiment of the present invention. FIG. 11 (a) is a photograph of a slitting yarn which is firstly slit with a width of 50 mm, and FIG. 11 11 (b) is a photograph showing the process of precisely slitting the primary slit yarn with a width of 1.5 mm, and FIG. 11 (c) is a photograph showing the process of slitting the primary slit yarn with a width of 1.5 mm A photo showing the process of becoming,

FIG. 12 is a view showing an image of a cell aggregate specimen cultured in a slitting furnace included in an embodiment of the present invention, taken along a confocal microscope at the height from the lower side to the upper side of the specimen in a state in which the cell aggregate is attached, and

13 is a SEM photograph of a first yarn performing a support function included in an embodiment of the present invention.

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings, which will be readily apparent to those skilled in the art to which the present invention pertains. The present invention may be embodied in many different forms and is not limited to the embodiments described herein. In order to clearly illustrate the present invention, parts not related to the description are omitted, and the same reference numerals are assigned to the same or similar components throughout the specification.

First, the horny yarn in the first embodiment according to the present invention will be described.

As shown in FIGS. 1 to 4, the horny sheaths 10, 10 ', 20, and 100 according to an embodiment of the present invention have amino acid sequences in which the first motif and / or the first motif amino acid sequence are repeated at least twice the first motif conjugates _(1b, 21a _2, 31b) first yarns (1, 1 ', 21,30) and said first motif, and / or wherein the first conjugate motif _(1b, 21a _2, 31b) containing the (2, 2 ', 22, 41) comprising a second motif combination (2b, 22a2, 41b) having an amino acid sequence in which the heterologous second motif and / or second motif amino acid sequence is repeated at least twice, Respectively.

The heterogeneous yarns 10, 10 ', 20, and 100 according to the present invention may include motifs that are as heterogeneous as the number of yarns constituting the heterogeneous yarn. However, heterogeneous motifs may be spatially separated by including heterogeneous motifs per yarn. As a result, the cells attached to the surface of any one of the yarns can be stimulated only by motifs that perform a specific kind or a specific function of the yarn, and can be stimulated by other motifs So that each cell contained in a cell population cultured in any one of the yarns may have similar characteristics or the same differentiation stage.

In addition, the function of each motif can be fully expressed as heterogeneous motifs do not physically and / or chemically combine with each other. That is, if two or more motifs, each of which is heterogeneous, are provided in one yarn, a solution obtained by mixing the first motif and the second motif may be treated on the yarn, or the first motif may be treated and then the second motif may be treated on the yarn These methods can cause coagulation between motifs in the manufacturing process, but the occurrence of chemical bonding can not be avoided. In this case, each of the motifs provided by the modification of the conformation for performing the original function of the motif or the occurrence of the portion of the amino acid sequence which causes the function of the original motif to appear due to the other motif, There is a problem that can not be performed completely. Accordingly, when the first motif is provided on one or more strands of the first yarn and the second motif is provided on one or more strands of the second yarn and then the yarns are mixed, even if different motifs are included in the mixed yarn, There is an advantage that the function can be fully manifested.

Accordingly, the cell culture supports 10, 10 ', 20, and 100 according to the present invention are characterized in that (1) the first motif and the first motif conjugate having an amino acid sequence in which the first motif amino acid sequence is repeated at least twice, And a second yarn having any one or more of the first motif, the second motif, and the second motif combination having an amino acid sequence in which the second motif and the second motif amino acid sequence are repeated at least twice, Preparing; And (2) concatenating the prepared first yarn and the second yarn.

First, in step (1) according to the present invention, a first yarn (1, 1 ', 21, 22) having any one or more of a first motif and a first motif combination having an amino acid sequence in which at least two first motifs are repeated, And a second motif (2, 2 ', 22, 41) having at least one of a second motif and a second motif combination having an amino acid sequence in which at least two second motifs are repeated with the first motif ), Respectively.

The motifs 1b, 21a ₂ , 31b, 2b, 22a ₂ and 41b provided in the first yarns 1, 1 ', 21 and 30 and the second yarns 2, 2' It may be one having a function of inducing adhesion, migration, growth, proliferation, and differentiation on the outer surface of the horn island of a cell to be treated, and in the case of a known motif performing such function, Can be employed. For example, the motif may be a natural peptide or a recombinant peptide comprising a predetermined amino acid sequence contained in at least one selected from a protein, a glycoprotein, and a proteoglycan contained in a growth factor or an extracellular matrix. . Specifically, the motif is selected from the group consisting of adrenomedullin, angiopoietin, bone morphogenetic protein (BMP), brain derived neurotrophic factor (BDNF), epidermal growth factor (EGF), erythropoietin, (GDNF), granulocyte colony-stimulating factor (G-CSF), granulocyte macrophage colony-stimulating factor (GM-CSF), fibroblast growth factor, glial cell line derived neurotrophic factor Growth differentiation factor-9 (GDF9), hepatocyte growth factor (HGF), hepatoma-derived growth factor (HDGF), insulin-like growth factor (IGF), keratinocyte growth factor (KGF), migration-stimulating factor (MSF), myostatin (GDF-8), nerve growth factor Platelet-derived growth factor cell growth factor (TCGF), neurofilin, transforming growth factor-alpha (TGF-a), transforming growth factor- (TGF-beta), tumor necrosis factor-alpha (TNF-alpha), vascular endothelial growth factor (VEGF), IL-1, IL-2, IL-3, IL-4, IL-5, IL-7. ≪ RTI ID = 0.0 > (GF) < / RTI > Or at least one extracellular matrix selected from the group consisting of hyaluronic acid, heparin sulfate, chondroitin sulfate, termarine sulfate, keratan sulfate, alginate, fibrin, fibrinogen, collagen, elastin, fibronectin, bitonectin, carderine and laminin or a sequence of a predetermined amino acid included in an extracellular matrix. In addition, the motif may include a predetermined amino acid sequence included in the growth factor and a predetermined amino acid sequence included in the extracellular matrix. More preferably, the motif may include at least one selected from the group consisting of proteins comprising the amino acid sequences of SEQ ID Nos. 8 to 28 and fusion proteins of at least two of these proteins, It is not.

Meanwhile, the motifs 1b, 21a ₂ , 31b, 2b, 22a ₂ , and 41b may be integrally formed by covalently bonding an adhesive component to the support to improve cell adhesion. For example, when it is intended to provide a peptide or polypeptide involved in cell growth, it is preferable to covalently bond the protein-binding component directly to the N-terminal and / or C-terminal of the peptide or polypeptide or to introduce a heterologous peptide or polypeptide In this case, a peptide or a polypeptide involved in the growth of the cell can be adhered more firmly to the supporting fiber, and the desorption of the cell can be minimized.

For example, the protein as such an adhesive component may also include a specific domain or amino acid sequence of a known mussel protein or mussel protein. The domain or amino acid sequence may be obtained by immobilizing the cultured cells on a cell support in the initial stage to prevent the cells loaded on the culture solution from floating and / or to immobilize other peptides and / or polypeptides involved in cell proliferation on the supporting fibers So that it can perform the function of preventing desorption of the supporting fibers during the cell culture. The adhesive component may be used without limitation in the case of a known adhesive component which is normally biocompatible and does not cause cytotoxicity, but preferably the amino acid sequence of SEQ ID NO: 1 to SEQ ID NO: 7 is repeated 1 to 20 times A protein, and a protein fused with at least two of these proteins. In this way, the cytotoxicity is markedly reduced, the adhesion of the motif is excellent, and the adhesive component is dissolved in the culture solution There is an advantage that desorption of a motif and isolation of a cell can be prevented.

According to a preferred embodiment of the present invention, the first motifs 1b, 21a _{2, and} 31b provided in the first yarns 1, 1 ', 21, and 30 may be components that promote cell proliferation and growth , And the second motifs (2b, 22a2, 41b) provided in the second yarns (2,2 ', 22,41) may be components that promote cell differentiation. The cells cultured in the embryonic stem thus cultured are more smoothly cultured and propagated in the first yarn, and when the cultured cells migrate to the second yarn, they are differentiated into specific stages through the differentiation promoting component provided in the second yarn It may be more advantageous to form the cell aggregate on the outer surface of the second yarn.

The motifs 1b, 21a ₂ , 31b, 2b, 22a ₂ and 41b are fixed to the outer surfaces of the first yarns 1, 1 ', 21 and 30 and the second yarns 2, For example, the components may be provided on the outer surface of the first yarn 1, 1 ', 21, 30 and the second yarn 2, 2', 22, 41 through a coating process . Alternatively, the motifs 1b, 21a ₂ , 31b, 2b, 22a ₂ , and 41b may be mixed with a fiber forming component together with a spinning liquid for producing a yarn and may be already provided in the state of yarn. In this case, there is an advantage that the motif can be easily provided on the outer surface of the produced yarn without a separate coating step or adhesive component.

As an example of providing the first motif in the first yarn, a coating method will be described in detail. A first motif solution in which the first motif is dissolved in a solvent is prepared. In view of the kind of the first motif, the solvent may be a known solvent which is excellent in solubility in the first motif but does not have the structural and chemical deformation of the first motif. For example, the solvent may be water or distilled water . At this time, the concentration of the first motif solution may be 0.01 to 0.10 mg / ml. If the concentration of the first motif is less than 0.01 mg / ml, the effect of the prepared motif may be insignificant, There is a concern that the adhesion may be physically weakened. If it exceeds 0.10 mg / ml, the content of the first motif coated may be insignificantly increased, aggregation between the first motifs may be excessive, There is a concern that the coated portion of the first motif may be separated by pipetting when the medium is exchanged because the adhesive strength may be physically weakened due to the content of the first motif. Next, the step of injecting EDC and NHS, which performs a function of adsorbing and coating the first motif on the surface of the first yarn, may be performed in the first motif solution. At this time, the EDC and NHS may be added to the first motif solution and then mixed to prepare a coating composition. Or an EDC solution in which EDC and NHS are respectively dissolved in a solvent and an NHS solution are mixed, the mixed solution is mixed in the first motif solution after 10 to 60 minutes and then reacted for 10 to 60 minutes to prepare a coating composition.

The prepared coating composition may then be treated with a coating method known to the first yarn, followed by washing and drying, and then the first yarn may be provided with the first motif. At this time, the first motif provided in the first yarn may be a first motif combined with one or more first motifs and / or several or several first motifs continuously.

Also, the second motif may be provided in the second yarn in the same manner as the above-mentioned method or may be suitably modified. In this case, the second motif may also be formed of one or more second motifs and / 2 < / RTI > motif combination.

Next, the first yarns (1, 1 ', 21, 30) and the second yarns (2, 2, 3) having the first motifs 1b, 21a _2, 31b and the second motifs 2b, 2 ', 22, 41) may be formed of a known fiber-forming component that can be produced in the form of fibers, and may be realized by selecting a suitable material according to the purpose, so that the present invention is not particularly limited thereto. The fiber forming component may include a natural fiber component such as a cellulose component such as cotton, hemp, etc., a protein component such as wool, silk, or a mineral component. Or the fiber forming component may be a component of known artificial fibers.

On the other hand, the fiber-forming component may contain, depending on the purpose, polystyrene (PS), polyester (e.g., polyethylene terephthalate), polyurethane sulfone (PES), polyvinylidene fluoride (PVDF), polyacrylonitrile (PDMS), polyamide, polyalkylene, poly (alkylene oxide), poly (amino acids), poly (allylamines), polyphosphazene ) And a polyethylene oxide-polypropylene oxide block copolymer, or a non-biodegradable component selected from the group consisting of polycaprolactone, polydioxanone, polyglycolic acid, PLLA a biodegradable component selected from the group consisting of poly (L-lactide), PLGA (poly-DL-lactide-co- glycolide), polylactic acid and polyvinyl alcohol .

The first yarn and the second yarn each independently including any one or more of the above-described fiber forming components may be formed of the same fiber forming component or may be formed of different fiber forming components.

The first yarns 1, 1 ', 21 and 30 and the second yarns 2, 2', 22 and 41 may be formed of one yarn or a yarn of yarn, yarn or slitting yarn, And may be included in a plurality of strands.

The spinning yarn may be produced through a raw cotton by a known method. Further, the filament yarn may be one produced by spinning by a known method, and the spinning may be a known spinning method such as chemical spinning or electrospinning.

The slitting yarns (21 and 22 in FIG. 3) may be manufactured by cutting a sheet-like fiber aggregate, a fabric or the like so as to have a predetermined width. Preferably, the slitting yarns may be made by cutting a sheet-like fibrous web having a three-dimensional network structure to have a predetermined width. At this time, the fibrous web may be pressed at a constant pressure to improve the ease of the slitting process and increase the strength of the slitting yarn. For example, when the sheet-like nanofiber web having the three-dimensional network structure of FIG. 5A is cut and cut to a predetermined width, a slitting yarn as shown in FIG. 5B can be manufactured. When a slitting yarn formed of a nanofiber web is provided as a yarn or a yarn, the culture solution can move between the outside and the inside of the yarn, thereby allowing cells to be cultured more smoothly. In addition, since the cells can propagate not only on the surface of the nanofiber web but also inside, the implementation of a three-dimensional cell cluster can be more advantageous.

However, even in the case of a nanofiber web designed to have many channels, it is difficult to pass the culture solution to a desired level because the flow path is reduced and the average pore size is reduced when compressed to be realized as a slitting yarn. In this case, it may be difficult to cultivate three-dimensional cell aggregates. Accordingly, the basis weight of the nanofiber web of the slitting yarn may be 0.1 to 100 g / m 2, more preferably 3 to 50 g / m 2, and still more preferably 7 to 30 g / m 2. If the basis weight is less than 0.1 g / m 2, the handling property is deteriorated, the slitting process tends to be unstable and unstable, and there is a fear of a decrease in productivity due to frequent trimming. If the basis weight exceeds 100 g / m < 2 >, the pressure of the nanofiber web is so large that the pores and the flow path inside the nanofiber web decrease significantly, and the sectional shape of the supporting fiber forming the nanofiber web is shaped like a pressed ellipse And in this case, there is a problem that the morphology of the nanofiber web surface or the inner void space is changed, making it difficult for the cells to grow in three dimensions and grow in a three-dimensional manner.

The thickness of the slitting yarn may be 7 to 30 탆, more preferably 9 to 25 탆. When the thickness of the slitting yarn 200 is thinner than 7 mu m, the handling property is deteriorated. In the twisting process performed after singly or several strands are folded together with a weak mechanical strength, threading may occur, . In addition, if the compression process is excessive and the thickness is reduced, the porosity and the average pore size may be reduced, and the surface morphology may become flat, making it difficult to stably cultivate the cells. If the thickness is more than 30 탆, it may be considered depending on the degree of basis weight. However, if the compression is less, the surface morphology may not be stable and cell desorption may occur frequently during culturing. In the case of thick cells, the porosity and the average pore size may be reduced. As a result, the three-dimensional culture of the cells may be difficult due to the decrease of the permeability of the cell culture medium and the decrease of the internal penetration of the cells.

The basis weight ratio (basis weight (g / m 2) ÷ thickness (μm)) to the thickness of the slitting yarn may be 1: 0.8 to 1.7, more preferably 1: 1 to 1.25, It is possible to cultivate a three-dimensional cell cluster more stably due to easy penetration of the inside of the tincture, and the permeation of the cell culture liquid is smooth, thereby minimizing the death of the cells loaded therein. If the basis weight ratio to the thickness is less than 0.8, the surface morphology may not be stable and cell desorption may be frequent during the culture. If the thickness is thick enough in a sufficiently compressed state, porosity and average pore size may be reduced. 3-dimensional culture of cells may be difficult due to decrease of permeability of cell culture fluid, decrease of internal penetration of cells, and the like. If the ratio of the basis weight to the thickness exceeds 1.7, the compression process may be excessively carried out, resulting in a thinner thickness, which may result in a decrease in porosity and average pore size, and a decrease in surface morphology And it may be difficult to stably cultivate the cells.

The slitting yarn may have a width of 0.1 to 8 mm. If the width of the slitting yarn is less than 0.1 mm, cutting is not easy and frequent cutting may be caused. Further, there is a problem in that it can be easily broken by the tension and the rotational force applied when the plurality of slitting yarns are twisted. Further, when the width is more than 30 mm, it may be difficult to attain the desired effect such that uneven twisting may occur in the continuous yarn manufacturing process in which a plurality of slitting yarns are joined together to produce a yarn.

In addition, the average pore size of the nanofiber web forming the slitting yarn is preferably such that the pores of the nanofiber web are formed so as to secure a space for the cells to move and propagate therein, and the diameter is determined according to the specific type of the cells to be cultured . For example, the average pore size may preferably be 0.05 to 10 mu m, more preferably 10 nm to 1 mu m. If the average pore diameter is less than 0.05 탆, the cells can be moved and propagated in a planar manner through the outer surface of the nanofiber web rather than moving into the pores inside the nanofiber web when the cultured cells proliferate, It may not be possible to cultivate the aggregate. In addition, even if the cells migrate into the inner space of the nanofiber web, the culture solution may not pass smoothly through the nanofiber web, resulting in the problem that the cells migrated to the inner space may be killed or the proliferation may be deteriorated. In addition, if the average pore size exceeds 10 탆, the cells may migrate into the internal space of the compressed nanofiber web and the culture solution may pass through, but the cells to be cultured may be mixed with the culture solution passing through the pressed nanofiber web The phenomenon of separation from the slitting yarn may be increased, and there is a problem that the increase in the isolated cultured cells is difficult to be cultured into a cell aggregate having the desired three-dimensional shape.

The average diameter of the supporting fibers forming the nanofiber web may be in the range of 10 nm to 100 mu m, preferably 100 nm to 50 mu m, more preferably 100 nm to 10 mu m, May be 100 nm to 1 占 퐉. If the average diameter of the supporting fibers is less than 10 nm, the mechanical strength of the fiber web may be significantly deteriorated. If the average diameter exceeds 100 탆, it may be difficult to produce a nanofiber web having the desired level of porosity and specific surface area .

On the other hand, in order to cultivate the cells in a three-dimensional manner, the cells penetrate and cultivate not only on the surface of the nanofiber web but also in the inner space, and eventually contact the cells penetrated and cultured in the inner space and the cells cultured on the surface, Cell aggregates can be formed. However, even in the case of the cells cultured on the surface, the three-dimensional culture of the cells can be induced by the surface morphology of the nanofiber web. As an example of this, the morphology of the nanofiber web surface realized by the slitting yarn is not flat, and the surface roughness may be large so as to be rugged. The ruggedness of the surface shape of the nanofibrous web includes, for example, a plurality of recesses and / or protrusions as examples. In addition to the effect of cubic growth of the cells, the cells are more easily and firmly adhered to the spaces between the protrusions or the grooves of the recesses There is an advantage that the number of cells to be desorbed after loading into the slitting yarn can be remarkably reduced.

According to a preferred embodiment of the present invention, the nanofiber web of the slitting yarn includes a plurality of supporting fibers in order to realize a yarn having a surface of a morphology for the three-dimensional growth of the cells and the improvement of the seating stability of seeded cells And the diameter dispersion coefficient (E) of the supporting fibers may be from 8 to 25%. The diameter dispersion factor (E) is a parameter that can be used to determine how close or wide the supporting fibers are in relation to the predetermined average diameter calculated in the distribution diagram based on the diameters of the supporting fibers. .

[Equation 1]

The meaning that the diameter dispersion coefficient (%) according to Equation (1) is 0% means that the standard deviation is zero, which means that the diameters of the plurality of supporting fibers included in the fiber web are all equal to the average diameter. Thus, on the contrary, increasing the number of diameter dispersive means that the number of supporting fibers having larger diameter and / or smaller diameter than the average diameter among the plurality of supporting fibers contained in the fibrous web increases.

The support according to an embodiment of the present invention may have a structure in which a plurality of concave portions and / or convex portions are disposed as described above, as the support member satisfies a predetermined average diameter and satisfies a dispersion factor of 8 to 25% It may be easier to implement a nanofiber web having a rugged surface morphology. However, when the number of dispersed phases is excessively large, the increase in the basis weight relative to the thickness may be large, which may result in a significant decrease in the average pore size and air permeability, and difficulty in flowing into the compressed nanofiber web of the cell culture liquid, Cell culture may be difficult within the nanofiber web, thus reducing cell culture efficiency. If the dispersion factor for diameter is less than 8%, uniformity of diameter of supporting fibers increases, smooth surface morphology is advantageous and uniformity of pore diameter is also increased, but seeded cells grow on surface rather than three- Or if the mean diameter of the supporting fibers is large, a pore structure having a large average pore size may be generated and the seeded cells may be desorbed, so that a desired level of three-dimensional cell aggregates may not be cultured. In addition, if the number of dispersions with respect to the diameter exceeds 25%, the average pore diameter of the nanofiber web becomes very small as the non-uniformity of the support diameter increases in a nanofiber web having a somewhat smaller average diameter, It may become difficult to introduce or exchange into the inside of the web, and the cells may be cultured along the surface rather than three-dimensionally cultured.

Further, the air permeability of the nanofiber web may be 1 to 10 cfm. One of the important factors in the three-dimensional growth of cells in the slitting yarn, which is one example of the first yarn or the second yarn, is whether or not it is capable of continuously supplying the materials necessary for cell culture. In the case of the cells arranged adjacent to the surface of the slitting yarn when the cells are three-dimensionally grown on the surface of the slitting yarn or the cells which are settled in the inner space of the slitting yarn and are cultured, As shown in Fig. Therefore, the air permeability of the nanofiber web may be 1 to 10 cfm for smooth inflow and outflow of the cell culture liquid into the compressed nanofiber web. If the air permeability is less than 1.0 cfm, the cells may be difficult to penetrate into the support, and the components that can dissolve the cell culture fluid or the biodegradable component may not pass smoothly. If the air permeability exceeds 10 cfm, a fiber web having a significantly low mechanical strength of the nanofiber web or a fineness of the fineness of the supporting fiber can be realized because it is realized as a slitting yarn, And it may be difficult to apply to a limited small volume incubator, and it may be difficult to cultivate in the desired amount due to desorption of seeded cells.

In addition, the nanofiber web may have a porosity of 40 to 90%, thereby facilitating formation of a three-dimensional cell aggregate through the cells migrating and propagating inside the nanofiber web, There is an advantage that the culture solution can be carried or the permeability of the culture solution can be improved. If the porosity is less than 40%, it is difficult to form a cell aggregate in a cubic phase, and there is a problem that cell migration and proliferation inside the nanofiber web may be killed. In addition, if the porosity exceeds 90%, there is a problem that the support may collapse during cell culture due to the weak mechanical strength of the support.

The first yarns 1, 1 ', 21, 30 and the second yarns 2, 2', 22, 41 may independently have a fineness of 0.01 to 300 denier. However, the present invention is not limited thereto and may be modified to suit the kind, size, shape and size of the cell cluster to be cultured.

Next, in step (2) according to the present invention, the step of juxtaposing the prepared first yarn and the second yarn is performed.

The yarn may be produced by separately spinning the first yarn 1, 1 ', 21, 30 and the second yarn 2, 2', 22, 41 by a known yarn joining method such as air jet, So that the filament yarn can be manufactured in a known structure and shape. 1, 2 and 4, the first yarns 1, 21, 30 and the second yarns 2, 22, 40 are twisted to provide a three-dimensional growth space and a movement path of the cells 20, and 100 in which a plurality of kites are formed. Alternatively, as shown in FIG. 3, the first yarn 1 'and the second yarn 2' may have a twisted yarn (not shown) for preventing density-dependent inhibition of cultured cells after the twisted yarn and improving the contact- May be embodied as a horny yarn 10 'in which at least a part of the first yarn 1' and the second yarn 2 'of the yarn are merged to form a space separated from one yarn.

However, the present invention is not limited to the shapes shown in Figs. 1 to 4, and it is also possible that either one of the first yarn and the second yarn is judged and the other yarn is covered by the covering, The arrangement of the first yarn and the second yarn in the mixed yarn such that the first yarn and the second yarn cover the third yarn on the yarn may be changed according to the purpose.

The mixed yarn according to one embodiment of the present invention implemented through the above-described manufacturing method includes a first yarn having a first motif and a second yarn having a second motif different from the first motif.

The mixed yarn according to an embodiment of the present invention includes at least two motifs, but the first motif and the second motif are processed into a single mixed yarn in which the first yarn and the second yarn are mixed with each other The first yarn or the second yarn may not include the at least one first motif and the at least one second motif in a chemically or physically bonded form, and may not include the first motif and the second motif, There is an advantage that the function of each of the second motifs can be fully expressed. The presence or absence of a physical / chemical bond between the first motif and the second motif can be qualitatively and quantitatively analyzed using conventional protein detection methods such as TBO staining, Silver staining, and SDS page electrophoresis.

Next, the horny yarn according to the second embodiment of the present invention will be described.

5, the horny sheath 200 for a cell culture support according to an exemplary embodiment of the present invention includes a first yarn 220 having a supporting function and a second yarn 220 having a supporting function and at least a part of the outer surface of the first yarn 220 And a second yarn 210 to be coated. The second yarn 210 includes nanofibers 211 having an average diameter of 1000 nm or less.

5, the outer circumference of one strand 220 is covered with one strand 210, but the present invention is not limited thereto. As shown in FIG. 2, the two strands 210 may include two or more strands 210, May be coated on the outer surface of the second layer 220 '. Further, it is noted that the second yarn can cover not only one strand but also a plurality of strands of one strand or a plurality of strands of the first strand.

At this time, the covering may be realized by covering the outer surface of the first yarn by winding the second yarn, but the present invention is not limited thereto. The second yarn 210 is coated on the entire surface of the first yarn 220 except for the front end and the end of the first yarn 220. However, the second yarn 210 may be coated on the entire surface including the front end and the end of the first yarn 220, Or only a portion of the first yarn 220 may be covered.

On the other hand, when cells are cultured on a support formed of nanofibers considering the size of the cells, it is difficult for the cells to stably adhere to the nanofibers, and even if adhered to the supports, the fluid is easily separated from the support due to fluid flow and peripheral vibrations, The culture and growth rate may be lowered. Also, the cells adhering to the surface of the nanofibers due to the flow of the nanofibers can form a nonuniform aggregate, and it is difficult to differentiate and recover the cells. Furthermore, in order to efficiently cultivate the cells, in the spinning solution containing the functional material together with the spinning solution for the production of nanofibers, there is a possibility that the mechanical strength is lowered due to the compatibility problem between the functional material and the component forming the nanofiber The nanofibers may be refracted during the cell culture or the support formed including the nanofibers may collapse, so that it may not be possible to provide a stable environment as a support for cell culture. In addition, when a fabric that is woven or knitted using the non-nano fiber itself is used as a cell culture support, there is a problem that it is difficult to retain mechanical strength enough to perform a weaving or knitting process.

Accordingly, in order to compensate the mechanical strength of the second yarn 210, which is a nanofiber, the first yarn 220 having a supporting function is used as a second yarn 210 for the cell culture support according to the second embodiment of the present invention, The second yarn 210 is disposed outside the first yarn 220 so that the first yarn 220 is core yarn and the second yarn 210 is aligned with the second yarn 210, By making it possible to contact and cultivate the yarn 210, it is possible to stably cultivate cells while improving the efficiency of cell culture.

In addition, through this arrangement, the degree of adhesion of the second yarn 210 can be increased to prevent slip and improve tensile strength and durability. That is, although the second yarn 210 is composed of nanofibers considering the size of cells according to an example of the present invention, the second yarn 210 is twisted or coated on the first yarn 220, So that the cutting rate or the breakage rate of the second yarn 210 can be significantly lowered.

The first yarn 220 is a circular yarn that performs the supporting function by securing the mechanical strength of the second yarn 210 including the nanofibers specifically described below.

The first yarn 220 may be a yarn having a conventionally known fiber material, and as a non-limiting example, the fiber forming component may be a polystyrene (PS), a polyester, a polyether sulfone (PES), a polyvinyl (PVDF), polyacrylonitrile (PAN), polydimethylsiloxane (PDMS), polyamide, polyalkylene, polyalkylene oxide, polyurethane, poly amino acid (s)), poly (allylamines), polyphosphazene, and polyethylene oxide-polypropylene oxide block copolymers, or polycaprolactone, Polyglycolic acid, PLLA (poly-L-lactide), PLGA (poly-DL-lactide-co-glycolide), polylactic acid and polyvinyl Made of polyvinyl alcohol And at least one biodegradable component selected from the group consisting of

Also, as shown in FIGS. 5 and 6, the first yarns 220 and 220 'may be a single yarn having a single yarn as shown in FIG. 1, or a multi yarn having a plurality of mono yarns as shown in FIG. At this time, the first yarn can be multi-yarn in order to have a more improved specific surface area and surface morphology, and more preferably, the second yarn covering the outside of the first yarn can be multi-yarn, more preferably, It may be a form having a space that is separated from the monolithic body and partially separated. In this case, the curvature of the outer surface of the first yarns 220'A and 220'B significantly increases, or the cells that are cultured due to the increase in the bending that can be expressed by the gap between the monosas with the curvature of the outer surface And the culture efficiency can be remarkably improved.

In addition, the first yarns 220 and 220 'may have an average diameter of 100 to 1000 mu m. In this case, when the first yarn is a multi-yarn including a plurality of mono yarns, the average diameter of the mono yarns may be 50 to 500 mu m and the number of strands may be 2 to 15, but the present invention is not limited thereto.

Next, the second yarn 210 covering the outside of the first yarns 220, 220 ', 220'A, 220'B will be described.

The diameter of the second yarn 210 may be determined in consideration of the type and size of the cells to be cultured. However, the diameter of the second yarn 210 may be determined by considering the size of the nano- Fibers, preferably 100 to 900 nm. If the average diameter of the nanofibers exceeds 1000 nm, it may be difficult to produce a three-dimensional cell aggregate at a desired level due to a decrease in the specific surface area to which the cells adhere, or the cell culture rate may be lowered. On the other hand, the average diameter of the nanofibers may be 100 nm or more. If the average diameter is less than 100 nm, the yarn is frequently stuck at the time of manufacturing the second yarn, resulting in lower productivity, It may be difficult to arrange the yarn, and it may be difficult to stably cultivate the cells even when the cells are cultured.

In addition, the second yarn may be a known fiber-forming component that can be manufactured using nanofibers, and the material may be selected in consideration of a predetermined purpose such as decomposability. Accordingly, And is not particularly limited. The fiber forming component may include a natural fiber component such as a cellulose component such as cotton, hemp, etc., a protein component such as wool, silk, or a mineral component. Or the fiber forming component 110 may be a component of known artificial fibers.

On the other hand, the fiber forming component forming the second yarn may be selected from the group consisting of polystyrene (PS), polyester, polyether sulfone (PES), polyvinylidene fluoride (PVDF), polyacrylonitrile (PDMS), polyamides, polyalkenes, poly (alkylene oxides), polyurethanes, poly (amino acids), poly (allylamines), polyphosphazenes polyglycolic acid, polylactic acid, polyglycolic acid, polylactic acid, polyglycolic acid, polyglycolic acid, polylactic acid, polyglycolic acid, polyglycolic acid, , at least one biodegradable component selected from the group consisting of poly (L-lactide), PLGA (poly (DL-lactide-co- glycolide)), polylactic acid and polyvinyl alcohol include Can.

Referring to FIG. 5, the second yarn 210 may be a slitting yarn in which a nanofiber web having a three-dimensional network structure formed of nanofibers 211 is cut to a predetermined width, and the second yarn 210, The horn yarn 200 may be embodied by covering the outer surface of the first yarn 220. At this time, the slitting yarn may be provided as one strand or a plurality of strands to form a second yarn.

Since the slitting yarn is the same as the description of the slitting yarn described in the first embodiment, detailed description thereof will be omitted.

On the other hand, when the slitting yarn is a second yarn, the first yarn and the second yarn are mixed at a weight ratio of 1: 0.5 to 10, more preferably 1: 1 to 5, more preferably 1: 1 to 2 . If the second yarn is less than 0.5 weight ratio as compared to the weight of the first yarn, it may be difficult to realize the three-dimensional cell aggregation by the reduction of the specific surface area of the mixed yarn. Also, if the second yarn is more than 10 weight ratio as compared to the weight of the first yarn, it may be difficult to improve the mechanical strength to a desired level, and when the cell culture fluid is subjected to flow or vibration, It may be difficult to stably cultivate and it may be difficult to weave or knit with a fabric.

In the case where the slitting yarn is the second yarn, the blended yarn may be twisted so that the second yarn 210 as the slitting yarn surrounds the first yarn 220. In this case, the surface area to which the cells can be attached is widened, so that the cells constituting the community can be easily cultured and the cells can be cultured at the same time. In this case, the extent to which the second yarn 210 is twisted with the first yarn 220 can be determined in consideration of the type, size, shape and size of the cell aggregate to be cultured. For example, the second yarn 210 may have a softening water of 10 to 100,000 T / cm. If the number of years is less than 10 T / cm, the second yarn is shaken due to an external force such as a flowing cell culture liquid, and if the second yarn has a plurality of strands of monosus, There is a problem that the cells may be difficult to be stably cultured. In addition, if the number of years exceeds 100,000 T / cm, the strength of the second yarn or the mixed yarn in which the second yarn and the first yarn are joined due to the friction between the mono yarns or the mono yarns may be lowered, The degree of adhesion between the soft tissue and the soft tissue may be severe and the volume of the fibrous bone may decrease.

According to another embodiment of the present invention, as shown in FIGS. 8 and 9A to 9C, the horn yarn yarn 300 includes a second yarn 310 directly wound on the outer surface of the first yarn 320, And the second yarn 310 may be a nanofiber aggregate formed of the electrospun nanofibers 311 and 312. In this case, In this case, the nanofibers of the three-dimensional network structure can be formed by direct-spun nanofibers folded and arranged / laminated a plurality of times without orientation. Or a nanofiber web having a three-dimensional network structure can be formed by arranging / stacking each of the nanofibers including a plurality of nanofibers independently without folding and / or fixing the length of the fibers. At this time, bonding may occur between the different surfaces and / or the surfaces of the different nanofibers in the one-stranded nanofibers, thereby making the three-dimensional network structure more structurally more complex. In this case, the cells loaded on the supporter can migrate / propagate into pores formed in a three-dimensional network structure, and may be advantageous for culturing cells with a cell aggregate having a three-dimensional shape / structure. In order to increase the proliferation rate and survival rate of the cells cultured inside / outside the support, it is important to supply the nutrients necessary for cell proliferation. In the nanofiber web of the three-dimensional network structure, As the channel is formed, it is possible to easily supply nutrients to the cells located inside the support, thereby preventing cell death and improving cell proliferation.

According to the amount of the nanofibers to be radiated, the nanofibers include nanofibers 311.312, a surface portion covering the outer surface of the first yarn, and a portion of the nanofibers extending outward from the surface portion. And may include protruding protrusions. At this time, since the surface portion is formed by the nanofibers 311 disposed in contact with the outside of the first yarn 320, the outer surface of the surface portion has a surface with many bends due to the outer surface of the nanofibers 311 Such curvature can increase the specific surface area of the support to enable smooth attachment and cultivation of cells, and can have a more suitable surface morphology for cell culture. In addition, the nanofibers 312 of the protrusions also increase the specific surface area of the second yarn and enable the implementation of various surface morphologies.

At this time, in order to improve the adhesion between the first yarns, a portion of the nanofibers 311 of the surface portion may be partially or entirely bonded to each other to cover the outside of the first yarn. If the individual nanofibers are provided on the surface of the first yarn without forming a surface portion in a predetermined region, it is difficult to have a sufficiently large specific surface area, and the individual nanofibers may be separated from each other by external force, that is, As the cells are easily shaken, the loaded cells may be difficult to stably cultivate and, in some cases, the cells may desorb from the support.

In the embodiment shown in FIG. 8, the first yarn may be provided as a multi yarn having a plurality of monos yarns as shown in FIGS. 9A and 9B, thereby enhancing the adhesion between the nanofiber aggregate and the first yarn And stable cell culture may be possible. More preferably, the first yarn may be partially twisted after the multi yarns are twisted as shown in FIG. 7B, and through this, a greater adhesion force can be exhibited by the increase of the specific surface area capable of joining with the nanofibers forming the nanofibrous aggregate There is an advantage to be able to do.

If the thickness exceeds 5 mu m, the surface morphology of the surface portion may be reduced, and the specific surface area may be decreased, so that the cell culture efficiency may be lowered. It may be difficult to cultivate a cell aggregate of a dimension. If the thickness is less than 0.2 탆, there is a fear that the adhesion with the first raw yarn is reduced, and the flow of the second yarn is intensified, so that it is difficult for the cells to be stably expressed. In addition, it may be difficult for the cells to migrate into the surface portion, which may make it difficult to cultivate into a three-dimensional cell cluster.

When the nanofiber aggregate is a second yarn, the first yarn and the second yarn are mixed in a weight ratio of 1: 2 to 5, more preferably 1: 1 to 3, more preferably 1: 1 to 2 As shown in FIG. If the amount of the second yarn is less than 2 parts by weight relative to the weight of the first yarn, it may be difficult to realize the three-dimensional cell aggregation by the reduction of the specific surface area of the mixed yarn. Also, if the second yarn is more than 5 wt% in comparison with the weight of the first yarn, the accumulation and bonding of the nanofibers on the outer surface of the horn sheath yarn may become severe and it may be difficult to sufficiently improve the specific surface area. It is difficult to move into the inside of the cell, and cultivation into a three-dimensional cell cluster may be difficult. In addition, the cell culture medium may be difficult to stably cultivate under the condition of flow or vibration, and it may be difficult to weave and knit as a fabric.

When the second yarn is a nanofiber aggregate, the hybrid fiber may be realized by electrospinning the nanofibers outside the first yarn. The electrospinning may include, for example, spinning the first yarn and radiating the nanofibers from the plurality of nozzles in one direction Or by radiating a nano-island in a plurality of nozzles fixedly arranged by dividing 360 ° at a predetermined angle around the first yarn, or moving the nozzle by 360 ° around the first yarn . The specific method is not limited thereto, and can be implemented through appropriate modification using the electrospinning device.

Meanwhile, the second yarns 210 and 310 may include a physiologically active component that induces at least one of cell adhesion, migration, growth, proliferation, and differentiation.

The physiologically active substance may be a substance which can aid in the cultivation of cells or can assist in the recovery of cultured cells.

The physiologically active component may be coated on the outer surface of the nanofibers included in the second yarn, or may be dispersed in the mixed state in the manufacturing step of the spinning solution for forming the nanofibers.

The physiologically active component may be, for example, an adhesive component, thereby fixing the cultured cells on the cell support in the early stage to prevent floating of the cells loaded on the culture solution and preventing the cells from being separated from the support during culture Can be performed. The adhesive component may be used without limitation in the case of a known adhesive component which is normally biocompatible and does not cause cytotoxicity, but preferably the amino acid sequence of SEQ ID NO: 1 to SEQ ID NO: 7 is repeated 1 to 20 times A protein, and a protein in which at least two of these proteins are fused. In this way, the cytotoxicity is remarkably reduced and the adhesive strength is excellent, and at the same time, the adhesive component is dissolved in the culture solution There is an advantage that desorption of the generated physiologically active ingredient and isolation of the cells can be prevented.

In addition, the physiologically active component may directly or indirectly induce cell adhesion, migration, growth, proliferation, and differentiation of cells so as to promote cell culture in addition to adhering cells to the support And may be included without limitation as long as it is a known substance known to express such a function. For example, the physiologically active ingredient may be a monoamine, an amino acid, a peptide, a saccharide, a lipid, a protein, a glucoprotein, a glucolipid, a proteoglycan, a mucopolysaccharide, acid, and a cell or a cell. The monoamines include, for example, compounds containing primary amines such as catecholamines and indole amines. Further, the peptide includes an oligopeptide, and the protein includes a polypeptide, such as, for example, fibronectin. The saccharides include monosaccharides, polysaccharides, oligosaccharides, and carbohydrates, and the lipids may be, for example, steroid hormones.

On the other hand, the physiologically active ingredient may include a motif. Since the description of the motif is the same as the description of the motif of the first embodiment, the description is omitted here.

In addition, the nanofiber forming the second yarn may further include a fiber cutting component. Generally, after cells are cultured, a process of separating the cells from the cultured supporter is necessary in order to transfer them to the target transplant. At this time, the cells that are cultured due to cell adhesion and clusterability can not be easily disappeared from the supporter, resulting in damage to the cell or damage to the supporter. In particular, in the case of the colony-forming cells characterized in that they are cohered with each other, the cells are not easily recovered, and thus a culture technique or a functional substance for separating from the cell culture support tissue is required.

Accordingly, the heterogeneous yarn 200 for the cell culture support according to the second embodiment of the present invention further includes a fiber cutting component in the second yarn 210 in order to easily recover the cells without damaging the cells cultured in the support can do. The fiber cleavage component may comprise a component for known cell recovery.

The fiber cutting component may be included in a spinning solution for implementing nanofibers. In this case, the fiber cutting component may be exposed on a part of the surface of the nanofiber, and some of the fibers may be disposed inside the nanofiber. have. The fiber-cutting component can cause the nanofiber itself to be cut off around the fiber-cutting component, so that the cell aggregate, which is a cultured product, can be easily separated from the filament yarn. For example, the fiber cutting component can impart a predetermined stimulus to the nanofiber and change the surface of the nanofiber to a non-adhesive state to separate the cultured cell and the hybrid fiber. Or the fiber cutting component is removed in the fiber by a method such as dissolution, so that the cell aggregate can be separated by directly cutting the nanofiber.

The present invention, which further comprises such a fiber cleavage component, enables the cells to be released from the supporter without damaging the cultured cell aggregate and the supporter, so that the cell recoverability can be increased.

Meanwhile, the present invention can be embodied as a fabric for a cell culture support comprising the hornblende yarn according to the present invention.

The fabric may be any one of a fabric, a knitted fabric, and a nonwoven fabric, and may be manufactured in different forms depending on the purpose. The fabrics, knit fabrics and nonwoven fabrics can be made through each of the known implementation methods. For example, the fabric may be a twill fabric woven by incorporating the above-mentioned hornblende yarn singly or in combination into at least one of warp and weft yarns. Further, for example, the knitted fabric may be a flat knitted fabric by putting the above-mentioned horn yarn yarn described above singly or in combination into a weft knitting machine. For example, the non-woven fabric may be a short-cut yarn obtained by cutting the horny filament yarn into a predetermined filament yarn by using heat / pressure with an adhesive component added thereto.

Also, a tissue engineering implant including cells cultured by implanting cultured cells into the above-described fabric according to the present invention can be implemented. At this time, the cultured cells can be cultured in a space between the first yarn of the mixed yarn and the space between the outer surface of the second yarn and the horny sheath.

At this time, the cell may be any one or more stem cells selected from the group consisting of ovarian stem cells, pluripotent stem cells, pluripotent stem cells, oligopotent stem cells, and single stem cells, and hematopoietic stem cells, hepatocytes, The present invention may include at least one of differentiation cells selected from the group consisting of cells, mesothelial cells, endothelial cells, muscle cells, nerve cells, immune cells, adipocytes, cartilage cells, bone cells, blood cells and skin cells. For example, the cell may be a cell having a shape elongated in one direction rather than a sphere, or a cell having strong mobility.

SEQ ID NO:	Amino acid sequence
One	Met Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys AlaLys Pro Ser Tyr Lys Ala Lys Pro Ser Tyr Pro ProThr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ser Ser GluGlu Tyr Lys Gly Gly Tyr Tyr Pro Gly Asn Thr Tyr His Tyr His Ser Gly Ser Ser Tyr His Gly Ser Gly Tyr His Gly Lys Tyr Lys Asn Ser GlyLys Tyr Lys Tyr Leu Lys Lys Ala Arg Lys Tyr His Arg Lys Gly TyrLys Lys Tyr Tyr Gly Gly Ser Ser Ala Lys Pro Ser Tyr Pro Pro ThrTyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro SerTyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lysla Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr ProPro Thr Tyr Lys
2	Met Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys AlaLys Pro Ser Tyr Lys Ala Lys Pro Ser Tyr Pro ProThr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ser Ser GluGlu Tyr Lys Gly Gly Tyr Tyr Pro Gly Asn Thr Tyr His Tyr His Ser Gly Ser Ser Tyr His Gly Ser Gly Tyr His Gly Lys Tyr Lys Asn Ser GlyLys Tyr Lys Tyr Leu Lys Lys Ala Arg Lys Tyr His Arg Lys Gly TyrLys Lys Tyr Tyr Gly Gly Ser Ser Ala Lys Pro Ser Tyr Pro Pro ThrTyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro SerTyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr LysAla Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr ProPro Thr Tyr Lys Gly Arg Gly Asp Ser Pro
3	Met Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys AlaLys Pro Ser Tyr Lys Ala Lys Pro Ser Tyr Pro ProThr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Pro Trp AlaAsp Tyr Tyr Gly Pro Lys Tyr Gly Pro Pro Arg Arg Tyr Gly Gly GlyAsn Tyr Asn Arg Tyr Gly Arg Arg Tyr Gly Gly Tyr Lys Gly Trp AsnAsn Gly Trp Lys Arg Gly Arg Trp Gly Arg Lys Tyr Tyr Gly Ser AlaLys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro ProThr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys ProSer Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr TyrLys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Leu
4	Ala Asp Tyr Tyr Gly Pro Lys Tyr Gly Pro Arg Arg Tyr Gly GlyGly Asn Tyr Asn Arg Tyr Gly Arg Arg Tyr Gly Gly Tyr Lys Gly TrpAsn Asn Gly Trp Lys Arg Gly Arg Trp Gly Arg Lys Tyr Tyr
5	Ser Ser Glu Glu Tyr Lys Gly Gly Tyr Tyr Pro Gly Asn Thr Tyr His Tyr His Ser Gly Gly Ser Tyr His Gly Ser Gly Tyr His Gly Gly Tyr Gly Lys Gly Lys Tyr Tyr Gly Lys Ala Lys Lys Tyr Tyr Tyr Lys Tyr LysAsn Ser Gly Lys Tyr Lys Tyr Leu Lys Lys Ala Arg Lys Tyr His ArgLys Gly Tyr Lys Lys Tyr Tyr Gly Gly Gly Ser Ser
6	Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys
7	Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys Ala LysPro Ser Tyr Pro A Pro Lys Pro Ser Tyr Lys Ala Lys Pro Ser Tyr Pro Pro ThrTyr Lys Ala Lys Pro Ser Tyr Pro Pro Thr Tyr Lys
8	Arg Gly Asp
9	Arg Gly Asp Ser
10	Arg Gly Asp Cys
11	Arg Gly Asp Val
12	Arg Gly Asp Ser Pro Ala Ser Ser Lys Pro
13	Gly Arg Gly Asp Ser
14	Gly Arg Gly Asp Thr Pro
15	Gly Arg Gly Asp Ser Pro
16	Gly Arg Gly Asp Ser Pro Cys
17	Tyr Arg Gly Asp Ser
18	Ser Pro Pro Arg Arg Ala Arg Val Thr
19	Trp Gln Pro Pro Arg Ala Arg Ile
20	Asn Arg Trp His Ser Ile Tyr Ile Thr Arg Phe Gly
21	Arg Lys Arg Leu Gln Val Gln Leu Ser Ile Arg Thr
22	Lys Ala Phe Asp Ile Thr Tyr Val Arg Leu Lys Phe
23	Ile Lys Val Ala Asn
24	Lys Lys Gln Arg Phe Arg His Arg Asn Arg Lys Gly Tyr Arg Ser Gln
25	Val Ala Glu Ile Asp Gly Ile Gly Leu
26	Pro His Ser Arg Asn Arg Gly Asp Ser Pro
27	Asn Arg Trp His Ser Ile Tyr Ile Thr Arg Phe Gly
28	Thr Trp Tyr Lys Ile Ala Phe Gln Arg Asn Arg Lys

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these examples are for illustrative purposes only and that the scope of the present invention is not construed as being limited by these examples.

≪ Example 1 >

First, to prepare a spinning solution, 12 g of polyvinylidene fluoride (Arkema, Kynar 761) as a fiber-forming component was added to 88 g of a mixed solvent of a mixture of dimethylacetamide and acetone at a weight ratio of 70:30 for 6 hours Using a magnetic bar to prepare a mixed solution. The prepared spinning solution was subjected to electrospinning in an environment of RH 65% and 30 ° C under the conditions of an applied voltage of 25 KV, a distance of 25 cm from the current collector to the spinneret, and a discharge amount of 0.05 ml / hole, A nanofiber web having a diameter of 402 nm, a thickness of 12.5 mu m, and a basis weight of 8 g / m 2 was obtained. The obtained nanofiber web was calendared twice at a temperature of 130 ° C and a pressure of 4 kPa to prepare a nanofiber web having a three-dimensional network structure, a basis weight of 10 g / m 2 and a thickness of 10 μm as shown in FIG. 10 (a) The manufactured nanofiber web was wound on a roll as shown in Fig. 11 (c) through a first cutting step with a width of 50 mm in Fig. 11 (a) and a precision cutting step with a width of 1.5 mm in Fig. 11 (b) The yarn was manufactured.

Meanwhile, SEQ ID NO: 18 in Table 1 was used as the first motif and SEQ ID NO: 20 in Table 1 was prepared as the second motif. Then, each of the motifs was dissolved in D.I. water to prepare a first motif solution and a second motif solution having a concentration of 0.1 mg / ml. In addition, 20 mM sodium acetate buffer, 1.52 g NHS and 1.33 g EDC were prepared. After that, NHS / EDC was dissolved in 350 ml of pH 6.5 20 mM sodium acetate buffer. After the treatment, the reaction was continued for 30 minutes. After that, the first motif solution and the NHS / EDC solution of the second motif solution were mixed and reacted for 40 minutes. After completion of the reaction, the solution was removed and washed three times using sterilized water. Work on the bench.

Thereafter, the slitting yarn is used as a first yarn and a second yarn, and the first coating composition is applied to the first yarn and the second coating composition is applied to the second yarn, respectively. After the washing process, And dried for 120 minutes to prepare a second yarn having a first yarn and a second motif provided with the first motif.

Then, the first yarn and the second yarn were twisted to prepare a filament yarn for a cell culture support as shown in Table 2 below.

≪ Comparative Example 1 &

The first motif solution prepared in the same manner as in Example 1 and the NHS / EDC prepared in the same manner as in Example 1, except that the coating solution for the first motif and the second motif were not separately prepared, Solution to prepare a single coating composition. The single coating composition thus prepared was treated and the dried first yarn and the second yarn were spun together to prepare a hornblend yarn for a cell culture support as shown in Table 2 below.

<Experimental Example 1>

The hornblende yarns prepared in Examples and Comparative Examples were cut to the same length and fixed on a cell culture well plate. Mesenchymal stem cells (MSC) were loaded at a density of 5 × 10 ⁴ cells / well in a well plate equipped with a cross-linker, and then grown in a 10% complete medium at 37 ° C. for 4 days. At this time, the 10% complete medium was prepared by mixing Ham's F12 medium in a modified eugles medium (DMEM) of Duvé course at a volume ratio of 1: 1.5, adding 7 vol% of fetal bovine serum, U / mL and streptomycin (65 mu g / mL) were added to the culture supernatant.

After incubation, the cultured mesenchymal stem cells (MSC) were stained with AP or Neutral red solution, and incubated for 10 minutes in an incubator. After staining with Cell Counting Kit 8 (CCK-8), a UV-vis spectrometer Was used to measure the absorbance. At this time, control was used in a cell culture dish which had been cultured in the same culture conditions.

The absorbances of Comparative Example 1 and Comparative Example 2 are shown in Table 2 based on 100% of the absorbance measured in Example 1 and Comparative Example 1,

The higher the absorbance, the better the cell culture after placement of the cells in the cell support.

	Example 1	Comparative Example 1
Relative absorbance (%)	100	84

As can be seen from Table 2, it can be seen that there is a significant difference in cell culture proliferation between Example 1 and Comparative Example 1, despite cell culture under the same conditions. Specifically, even in the case of Comparative Example 1, the cell culture was markedly reduced, even though both the first motif and the second motif for improving cell proliferation were included in the same manner as in Example 1, It can be expected that the improvement effect of the cell proliferation through the motif is hardly expressed as a result of the function decrease or incapability due to the combination of the motif and the second motif.

&Lt; Examples 2 to 7 >

The nanofiber web was prepared in the same manner as in the slitting yarn prepared in Example 1 except that the nanofiber web had a basis weight and thickness as shown in Table 3, Tongsa was used as a first yarn and a second yarn to prepare a horny filament for cell supports.

<Experimental Example 3>

The distribution of the diameters of the supporting fibers in the nanofiber web of slitting sieves used in the manufacturing processes of Examples 1 to 7 was measured.

The average diameter and the standard deviation of the diameter are calculated by using the distribution of the diameter of the supporting fibers measured by the method of the cyan distribution program (developed by AMOGREENTECH), and the average diameter and the dispersion coefficient are shown in Table 3.

<Experimental Example 4>

The air permeability of the nanofibrous webs of the slits used in the manufacturing processes of Examples 1 to 7 was measured and shown in Table 3 below.

The air permeability was measured by cutting the fiber web to a size of 38 cm2 using a TEXTEST instruments instrument and then grasping it with the equipment and measuring the amount of air passing through the air at a test pressure of 125 Pa. The unit of air permeability was cfm ft &lt; ³ &gt; / ft &lt; ² &gt; / min).

<Experimental Example 5>

The horny filaments prepared in Examples 1 to 7 were cut to the same length and fixed on a cell culture well plate. Fibroblasts (HS27) were loaded into the prepared well plates and then grown in 10% complete medium for 4 days at 37 ° C. At this time, the 10% complete medium was prepared by mixing Ham's F12 medium in a modified eugles medium (DMEM) of Duvé course at a volume ratio of 1: 1.5, adding 7 vol% of fetal bovine serum, U / mL and streptomycin 65 mu g / mL.

Thereafter, DAPI staining was performed on the proliferated fibroblasts, and the cells cultured through the Confocal microscope were attached. As a result, the height of the first source yarn, Photographs were taken and the number of cells cultured on the surface of the first source slitting yarn (the number of cells on the upper surface and the average number of cells on the lower surface) and the number of cells cultured in the first source, the slitting yarn And the average number of cells measured at 1/3 and 2/3 of the thickness) were measured and calculated. In Table 3, the number of surface cells and the number of internal cells measured in Example 1 were 100%, respectively, and the number of cells in each of the other examples was relatively shown. In addition, the measured photographs of the specimen section height according to Example 1 are shown in Fig.

Specifically, in FIG. 12, the cells cultured at a height of 7.6 μm from the lower side to the upper side of the specimen were observed, and it was confirmed that the cells were cultured to form a plurality of cell aggregates at a height of 15.3 μm. In addition, it can be expected that the height of the lower surface of the slitting yarn is between 15.3 mu m and 22.9 mu m considering that the nanofiber web formed of the cell aggregate cultured at a height of 22.9 mu m and the supporting fiber is observed. It was also confirmed that a large number of cell aggregates were cultured even at a height of 38.5 占 퐉 and a large amount of cell aggregates were cultured even at heights of 38.2 占 퐉 and 45.8 占 퐉 higher than the upper surface of the slitting yarn. It can be confirmed that the cells are uniformly grown three-dimensionally over the inside and the surface of the yarn.

	Example 1	Example 2	Example 3	Example 4	Example 5	Example 6	Example 7
Slitting Sine Nano Fiber Web Support Fiber Average Diameter (nm)	402	400	400	404	401	402	400
Dispersion coefficient of supporting fiber (%)	13.25	13.01	14.9	12.3	15.1	14.21	13.25
Basis weight (g / ㎡)	10	19	10	10	9.5	9	8
Thickness (㎛)	10	10.8	6.2	8	11.2	12.0	12.5
Basis weight (g / m < 2 >) / thickness (mu m)	One	1.76	1.61	1.25	0.85	0.75	0.64
Average pore size (nm)	500	430	458	472	519	538	565
Air permeability (cfm)	2.0	1.7	1.8	1.9	2.1	2.1	2.3
Number of surface cells (%)	100	75	87	95	91	76	58
Number of internal cells (%)	100	59	79	93	85	82	55

As can be seen from Table 3,

In the case of the slipping yarn of Example 7, it is structurally unstable, and the cells easily disappeared. As a result, it is difficult to stably cultivate the cells and the number of cultured cells is remarkably smaller than that of Example 1. [

In addition, among the examples, in the case of the mixed-fiber yarn including the slitting yarn according to Examples 1, 3, 4, and 5 having appropriate basis weight and thickness, the three- As a support for cell culture.

&Lt; Example 8 >

A PET multifilament yarn having a total diameter of 50 占 퐉 (monocular diameter of about 15 占 퐉) was prepared as a first yarn. Also, a slitting yarn was prepared as a second yarn. Specifically, to prepare a spinning solution, 12 g of polyvinylidene fluoride (Arkema, Kynar 761) as a fiber-forming component was added to 88 g of a mixed solvent in which a weight ratio of dimethylacetamide and acetone was 70:30, to 80 Lt; 0 &gt; C for 6 hours using a magnetic bar to prepare a mixed solution. The prepared spinning solution was subjected to electrospinning in an environment of RH 65% and 30 ° C under the conditions of an applied voltage of 25 KV, a distance of 25 cm from the current collector to the spinneret, and a discharge amount of 0.05 ml / hole, A nanofiber web having a diameter of 402 nm, a thickness of 12.5 mu m, and a basis weight of 8 g / m 2 was obtained. The obtained nanofiber web was calendared twice at a temperature of 130 ° C and a pressure of 4 kPa to prepare a nanofiber web having a three-dimensional network structure, a basis weight of 10 g / m 2 and a thickness of 10 μm as shown in FIG. 10 (a) The manufactured nanofiber web was wound on a roll as shown in Fig. 11 (c) through a first cutting step with a width of 50 mm in Fig. 11 (a) and a precision cutting step with a width of 1.5 mm in Fig. 11 (b) A second yarn was produced.

Then, the first yarn and the second yarn are twisted together so that the first yarn and the second yarn are in a weight ratio of 1: 2 so that the second yarn covers the outside of the first yarn, The same horny filaments were prepared.

&Lt; Examples 9 to 14 >

The nanofiber web was prepared in the same manner as the slitting yarn prepared in Example 8, and the nanofiber web was adjusted to have a basis weight and a thickness as shown in Table 2 by controlling the web weight / thickness of the nanofiber by varying the compression strength, Tonsa was used as the second yarn to prepare a hornblende yarn for a cell support as shown in Table 4 below.

&Lt; Comparative Example 2 &

Without the second yarn, the first yarn prepared in Example 8 was prepared as a yarn for cell culture support.

<Experimental Example 6>

The distribution of the diameter of the supporting fibers in the nanofiber web of the slitting sieve used in the manufacturing process according to Examples 8 to 14 was measured.

The average diameter and the standard deviation of the diameter are calculated by using the distribution of the diameter of the supporting fibers measured by the method using the cyan distribution program (developed by AMOGREENTECH), and the average diameter and the dispersion coefficient are shown in Table 4.

<Experimental Example 7>

The air permeability of the nanofiber webs of the slits used in the manufacturing processes of Examples 8 to 14 was measured and shown in Table 4 below.

The air permeability was measured by cutting the fiber web to a size of 38 cm2 using a TEXTEST instruments instrument and then grasping it with the equipment and measuring the amount of air passing through the air at a test pressure of 125 Pa. The unit of air permeability was cfm ft &lt; ³ &gt; / ft &lt; ² &gt; / min).

<Experimental Example 8>

The horns yarns prepared in Examples 8 to 14 and the first yarn according to Comparative Example 2 were cut to the same length and fixed on a cell culture well plate. Fibroblasts (HS27) were loaded into the prepared well plates and then grown in 10% complete medium for 4 days at 37 ° C. At this time, the 10% complete medium was prepared by mixing Ham's F12 medium in a modified eugles medium (DMEM) of Duvé course at a volume ratio of 1: 1.5, adding 7 vol% of fetal bovine serum, U / mL and streptomycin 65 mu g / mL.

Thereafter, DAPI staining was performed on the proliferated fibroblasts, and the cells cultured through the Confocal microscope were attached. From the lower part of the slitting yarn as the second origin of the horny filaments to the upper part, the height was varied in the Z- Photographs were taken and the number of cells cultured on the surface of the second source, the slitting yarn (the number of cells on the upper surface and the average number of cells on the lower surface) and the number of cells cultured in the second source, And the average number of cells measured at 1/3 and 2/3 of the thickness) were measured and calculated. In Table 4, the number of surface cells and the number of internal cells measured in Example 1 were 100%, respectively, and the number of cells in each of the other examples was relatively shown.

On the other hand, in the case of Comparative Example 2, photographs of the surface and the inside were taken at different heights in the Z-axis direction from the lower part to the upper part of the first yarn. As a result, it was confirmed that the cells were cultured only on the surface, Is shown as a relative value based on the number of surface cells of Example 1.

Specifically, in FIG. 8, cells cultured at a height of 7.6 μm from the lower side of the specimen to the upper side are observed, and it is confirmed that the cells are cultured to form a plurality of cell aggregates at a height of 15.3 μm. In addition, it can be expected that the height of the lower surface of the slitting yarn is between 15.3 mu m and 22.9 mu m considering that the nanofiber web formed of the cell aggregate cultured at a height of 22.9 mu m and the supporting fiber is observed. It was also confirmed that a large number of cell aggregates were cultured even at a height of 38.5 占 퐉 and a large amount of cell aggregates were cultured even at heights of 38.2 占 퐉 and 45.8 占 퐉 higher than the upper surface of the slitting yarn. It can be confirmed that the cells are uniformly grown three-dimensionally over the inside and the surface of the yarn.

	Example 8	Example 9	Example 10	Example 11	Example 12	Example 13	Example 14	Comparative Example 2
Slitting Sine Nano Fiber Web Support Fiber Average Diameter (nm)	402	400	400	404	401	402	400	Without second yarn
Dispersion coefficient of supporting fiber (%)	13.25	13.01	14.9	12.3	15.1	14.21	13.25
Basis weight (g / ㎡)	10	19	10	10	9.5	9	8
Thickness (㎛)	10	10.8	6.2	8	11.2	12.0	12.5
Basis weight (g / m < 2 >) / thickness (mu m)	One	1.76	1.61	1.25	0.85	0.75	0.64
Average pore size (nm)	500	430	458	472	519	538	565
Air permeability (cfm)	2.0	1.7	1.8	1.9	2.1	2.1	2.3
Number of surface cells (%)	100	73	85	93	90	78	53	25
Number of internal cells (%)	100	61	80	92	83	80	55	0

As can be seen from Table 4,

In the case of Comparative Example 2, as the second yarn was not included, the cells adhered and proliferated on the outer surface of the first yarn. However, it can be confirmed that the number of cells on the surface portion was significantly poor as compared with Example 1.

On the other hand, even in the case of using the second yarn, the slitting yarn of Example 14 is structurally unstable and the cells are easily separated, so that it is difficult to stably cultivate the cells and the number of cultured cells is remarkably smaller than that of Example 8 .

In addition, among the examples, a blended yarn including a slitting yarn according to Examples 8, 10, 11, and 12 having a basis weight and a proper thickness was prepared in the same manner as in Example 9, As a support for cell culture.

&Lt; Example 15 >

A PET multifilament yarn having a total diameter of 50 占 퐉 (monocular diameter of about 15 占 퐉) was prepared as a first yarn. In addition, the same spinning solution as in Example 1 was prepared in order to form the second yarn by directly spinning on the first source yarn through electrospinning.

After the prepared spinning solution was twisted, twelve nozzles were placed on the outer surface of the partially wound first yarn so as to be electrospun on the entire surface of 360 °, and electrospun to the first yarn so that the weight ratio was 1: 2.5 9a to 9c. &Lt; tb &gt; &lt; TABLE &gt;

&Lt; Example 16 >

The same procedure as in Example 15 was carried out except that the first yarn was twisted and then partially twisted without using any twist.

&Lt; Example 17 >

The same procedure as in Example 15 was carried out except that instead of the first yarn, a mono yarn constituting the first yarn was equally electrospun on the outer surface to provide a second yarn, Six strands were folded and twisted to form the filament yarns of Table 5 below.

<Experimental Example 7>

The horns yarns according to Examples 15 to 17 were fabricated and weighed to fabricate a fabric for a cell support having the same basis weight. The fabric was cut to a predetermined size and fixed on a cell culture well plate. Fibroblasts (HS27) were loaded into the prepared well plates and then grown in 10% complete medium for 4 days at 37 ° C. At this time, the 10% complete medium was prepared by mixing Ham's F12 medium in a modified eugles medium (DMEM) of Duvé course at a volume ratio of 1: 1.5, adding 7 vol% of fetal bovine serum, U / mL and streptomycin 65 mu g / mL.

Then, DAPI staining was performed on the proliferated fibroblasts, and the number of cells cultured on the surface of the fabric with attached cells attached to the confocal microscope was measured and calculated. Table 5 below shows the number of cells on the surface of the other examples relative to the number of surface cells measured in Example 15 as 100%.

<Experimental Example 8>

In order to evaluate the adhesion of the second yarn, the surface of the fabric implemented in Experimental Example 7 was photographed by an optical microscope to determine whether or not the nanofiber aggregate, which is the second yarn on the surface within the same area of the specimen according to Examples 15 to 17, Respectively. As a result of observation, the exposed area of the first yarn in the observation area was calculated as a percentage in the following Table 5.

	Example 15	Example 16	Example 17
Horn Temple	Uses first yarn as a multifilament yarn	Use yarn yarn as a multifilament yarn	After being electrospun in mono state,
Number of cells on the surface (%)	100	91	76
Exposure area of the outer surface of the first yarn (%)	9	15	42

As can be seen from Table 5,

It can be seen that Examples 15 and 16 using multi-yarn as the first yarn are much more excellent in adhesion of the second yarn than Example 17 and more suitable as a cell culture support.

Claims (14)

1. (1) a first yarn having at least one of a first motif and a first motif conjugate having an amino acid sequence in which a first motif amino acid sequence is repeated at least twice; and a second yarn having a second motif and a second motif, Preparing a second yarn each having at least one of a second motif binding substance having an amino acid sequence in which the motif amino acid sequence is repeated at least twice; And

   (2) folding the prepared first and second yarns together.
2. The method according to claim 1,

   Wherein the first yarn and the second yarn each independently comprise a spun yarn, a filament yarn, or a slitting yarn as one strand or a plurality of strands.
3. 3. The method of claim 2,

   (PS), polyester, polyether sulfone (PES), polyvinylidene fluoride (PVDF), polyacrylonitrile (PAN), polydimethylsiloxane (PDMS), polyamide, polyalkylene, polyalkylene But are not limited to, those selected from the group consisting of poly (alkylene oxide), poly (amino acids), poly (allylamines), polyphosphazene, and polyethylene oxide-polypropylene oxide block copolymers. One or more non-biodegradable components, or

   Polycaprolactone, polydioxanone, polyglycolic acid, PLLA (poly-L-lactide), PLGA (poly-DL-lactide-co-glycolide), polylactic acid Wherein the biodegradable component comprises at least one biodegradable component selected from the group consisting of polylactic acid and polyvinyl alcohol as a fiber forming component.
4. The method according to claim 1,

   Characterized in that the first yarn and the second yarn are partially melted after twisting or twisting.
5. The method according to claim 1,

   Wherein the first yarn or the second yarn of the mixed yarn does not include at least one first motif and at least one second motif in a chemically or physically bonded motif combination.
6. 3. The method of claim 2,

   Wherein the slitting yarn is a fibrous web of a three-dimensional network structure cut to have a predetermined width.
7. The method according to claim 6,

   Wherein the fibrous web is a nanofiber web having a basis weight to thickness ratio of 1: 0.8 to 1.7.
8. The method according to claim 6,

   Wherein the fibrous web is a nanofiber web having a basis weight of 5 to 30 g / m 2 and a thickness of 7 to 30 탆.
9. The method according to claim 1,

   Wherein the first motif and the second motif are motifs having a function of promoting at least one of attachment, migration, proliferation and differentiation of cells.
10. A first yarn having at least one of a first motif and a first motif combination having an amino acid sequence in which a first motif amino acid sequence is repeated at least twice; And

    And a second yarn having at least one of the first motif, the second motif, and a second motif conjugate having an amino acid sequence in which the second motif amino acid sequence is repeated at least twice.
11. 11. The method of claim 10,

    Wherein the first yarn or the second yarn of the mixed yarn does not include at least one first motif and at least one second motif in a chemically or physically bonded motif combination.
12. A fabric for a cell culture support comprising the hornblende yarn according to claim 11.
13. A fabric according to claim 12; And

    A tissue engineered graft containing the cells cultured in the fabric.
14. 14. The method of claim 13,

    The cell may be any one or more stem cells selected from the group consisting of ovarian stem cells, pluripotent stem cells, pluripotent stem cells, oligopotent stem cells, and single stem cells, and

    One of the differentiated cells selected from the group consisting of hematopoietic stem cells, hepatocytes, fibroblasts, epithelial cells, mesothelial cells, endothelial cells, muscle cells, nerve cells, immune cells, adipocytes, cartilage cells, bone cells, blood cells and skin cells Or more.

Images