CA2613408A1 - Time varying electromagnetic force sleeve for the expansion of cells and method of using the same - Google Patents
Time varying electromagnetic force sleeve for the expansion of cells and method of using the same Download PDFInfo
- Publication number
- CA2613408A1 CA2613408A1 CA002613408A CA2613408A CA2613408A1 CA 2613408 A1 CA2613408 A1 CA 2613408A1 CA 002613408 A CA002613408 A CA 002613408A CA 2613408 A CA2613408 A CA 2613408A CA 2613408 A1 CA2613408 A1 CA 2613408A1
- Authority
- CA
- Canada
- Prior art keywords
- electromagnetic force
- time varying
- varying electromagnetic
- electrically conductive
- force sleeve
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
- 238000000034 method Methods 0.000 title claims abstract description 32
- 230000010261 cell growth Effects 0.000 claims abstract description 10
- 210000004027 cell Anatomy 0.000 claims description 51
- 239000000203 mixture Substances 0.000 claims description 11
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims description 7
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 claims description 6
- 229910052802 copper Inorganic materials 0.000 claims description 6
- 239000010949 copper Substances 0.000 claims description 6
- 210000004976 peripheral blood cell Anatomy 0.000 claims description 5
- 239000004020 conductor Substances 0.000 claims description 4
- 239000004698 Polyethylene Substances 0.000 claims description 3
- 229910052742 iron Inorganic materials 0.000 claims description 3
- -1 polyethylene Polymers 0.000 claims description 3
- 229920000573 polyethylene Polymers 0.000 claims description 3
- 239000002738 chelating agent Substances 0.000 claims description 2
- 210000005259 peripheral blood Anatomy 0.000 claims description 2
- 239000011886 peripheral blood Substances 0.000 claims description 2
- 210000004504 adult stem cell Anatomy 0.000 claims 1
- 238000004113 cell culture Methods 0.000 description 8
- 238000012258 culturing Methods 0.000 description 5
- 230000012010 growth Effects 0.000 description 5
- 239000008055 phosphate buffer solution Substances 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- VVNCNSJFMMFHPL-VKHMYHEASA-N D-penicillamine Chemical compound CC(C)(S)[C@@H](N)C(O)=O VVNCNSJFMMFHPL-VKHMYHEASA-N 0.000 description 2
- 102000008100 Human Serum Albumin Human genes 0.000 description 2
- 108091006905 Human Serum Albumin Proteins 0.000 description 2
- 239000007760 Iscove's Modified Dulbecco's Medium Substances 0.000 description 2
- 239000003989 dielectric material Substances 0.000 description 2
- 230000005611 electricity Effects 0.000 description 2
- 239000004033 plastic Substances 0.000 description 2
- 229920003023 plastic Polymers 0.000 description 2
- 238000009827 uniform distribution Methods 0.000 description 2
- 230000000007 visual effect Effects 0.000 description 2
- 101000746367 Homo sapiens Granulocyte colony-stimulating factor Proteins 0.000 description 1
- 101000716729 Homo sapiens Kit ligand Proteins 0.000 description 1
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 1
- RTAQQCXQSZGOHL-UHFFFAOYSA-N Titanium Chemical compound [Ti] RTAQQCXQSZGOHL-UHFFFAOYSA-N 0.000 description 1
- 229910052770 Uranium Inorganic materials 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 230000004308 accommodation Effects 0.000 description 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 239000000919 ceramic Substances 0.000 description 1
- 229920001971 elastomer Polymers 0.000 description 1
- 230000005294 ferromagnetic effect Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 239000000122 growth hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 102000055151 human KITLG Human genes 0.000 description 1
- 239000012212 insulator Substances 0.000 description 1
- 239000011133 lead Substances 0.000 description 1
- 230000005291 magnetic effect Effects 0.000 description 1
- 239000000696 magnetic material Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229960001639 penicillamine Drugs 0.000 description 1
- 229920001296 polysiloxane Polymers 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000017423 tissue regeneration Effects 0.000 description 1
- 229910052719 titanium Inorganic materials 0.000 description 1
- 239000010936 titanium Substances 0.000 description 1
- JFALSRSLKYAFGM-UHFFFAOYSA-N uranium(0) Chemical compound [U] JFALSRSLKYAFGM-UHFFFAOYSA-N 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/02—Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M1/00—Apparatus for enzymology or microbiology
- C12M1/42—Apparatus for the treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
- C12M27/10—Rotating vessel
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N13/00—Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Sustainable Development (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Electromagnetism (AREA)
- Cell Biology (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
A time varying electromagnetic force sleeve wherein the time varying electromagnetic force sleeve comprises a time varying electromagnetic force source operatively connected to an electrically conductive coil that can removably receive a culture container. The present invention also relates to a method for cell expansion comprising providing a time varying electromagnetic force sleeve that is introduced to a culture container and that in use delivers a time varying electromagnetic force to cells contained within the culture container for cell expansion.
Description
TIME VARYING ELECTROMAGNETIC FORCE SLEEVE FOR THE
EXPANSION OF CELLS AND METHOD OF USING THE SAME
FIELD OF THE INVENTION
The present invention relates generally to a sleeve for the expansion of cells, and more particularly to a time va.rying electroinagnetic force sleeve that removably recieves a culture container. The present invention also relates to a time varying electromagnetic force sleeve that, when in use, supplies a time varying electromagnetic force to cells in a culture container for cell expansion therein.
BACKGROUND OF THE INVENTION
For many years there have existed methods for culturing cells. Some methods involve culturing cells in two-dimensional cultures utilizing such culture chambers as flasks and petri dishes, and others involve culturing cells in three-dimensional cultures utilizing such culture chambers as bioreactors. Methods of optimally culturing cells include adding molecules to cells in a culture such as growth factors, hormones, and others that, for instance, up or down regulate expansion of cells. Some methods are optimized for culturing individual cells and others are optimized for tissue culture. In addition, in an effort to produce more cells or larger tissue constructs over time, cell cultures are performed under various conditions.
United States Patent Number 6,673,597, Wolf et al., disclose the use of a time varying electromagnetic force ("TVEMF") to grow cells in culture. Wolf et al., disclose the use of electrodes to supply an electromagnetic force to a cell culture for growth of the same. In those instances where a conductive coil, rather than electrodes, is used for the delivery of a TVEMF to a cell culture, the coil is integral with a culture chamber and affixed thereto. Since the cells expanded in a culture may be introduced into the human body for tissue regeneration or treatment of human maladies, the culture chamber should be sterile. If the source of TVEMF to be delivered to cells in a culture chamber is integral with the culture chamber then either the culture chamber should be sterilized without disturbing the source of the TVEMF, a process which is cumbersome and time consuming, and if the container is to be discarded after each use, then the integral TVEMF coil has to be discarded therewith. Either way, an integral TVEMF coil is an expensive approach to cell expansion.
Consequently, it would be highly desirable to have a TVEMF sleeve comprising an electrically conductive coil having an interior portion wherein the interior portion defines a space wherein a culture container is removably received. It is also highly desirable to have a TVEMF sleeve comprising a coil support with an interior portion and an exterior portion, an electrically conductive coil wrapped around the exterior portion of the coil support; and a TVEMF source, and wherein the interior portion of the coil support defines a space wherein a culture container is removably received. It is fu.rther highly desirable to have a TVEMF sleeve that, when in use, is introduced to a culture container containing cells and supplies a TVEMF to the cells in a culture container for cell expansion. The present invention overcomes the problems associated with past and current cell culture methods, and presents advantages not before seen.
SUMMARY OF THE INVENTION
The present invention relates to a TVEMF sleeve comprising an electrically conductive coil having an interior portion and an exterior portion wherein the interior portion defines a space wherein a culture container is removably received, and a TVEMF
source operatively connected to the electrically conductive coil. Preferably the electrically conductive coil is substantially rigid.
The present invention also relates to a TVEMF sleeve comprising a coil support with an interior portion and an exterior portion wherein the interior portion defines a space in which a culture container is removably received, an electrically conductive coil wrapped around the exterior portion of the coil support, and a TVEMF source operatively connected to the electrically conductive coil.
The present invention also relates to a method of cell expansion comprising the steps of providing a time varying electromagnetic force sleeve comprising an electrically conductive coil, having an interior portion and exterior portion, operatively connected to a time varying electromagnetic force source; introducing a culture container containing cells to the interior portion of the electrically conductive coil of the time varying electromagnetic force sleeve; and supplying a time varying electromagnetic force to the cells in the culture container through the time varying electromagnetic force sleeve for cell expansion. The TVEMF sleeve may preferably deliver a TVEMF in the form of a pulsed square wave (following a Fourier curve) to the cells in the culture container, and the TVEMF may preferably be of from about 0.05 gauss to about 6 gauss.
Other aspects, features, and advantages of the present invention will be apparent from the following description of the preferred embodiments of the invention given for the purpose of disclosure. This invention may be more fully described by the preferred embodiment(s) as hereinafter described, but is not intended to be limited thereto.
BRIEF DESCRIPTION OF THE D.RAWINGS
In the drawings, Figure 1 is an elevated side view of a TVEMF sleeve;
Figure 2 is an elevated front view of a TVEMF sleeve;
Figure 3 is an elevated front view of a TVEMF sleeve removably adjacent to and encompassing a culture container;
Figure 4 is an elevated side view of a TVEMF sleeve;
Figure 5 is an elevated side view of a TVEMF sleeve being introduced to a culture container;
Figure 6 is an elevated side view of a TVEMF sleeve;
Figure 7 is a cross sectional elevated side view of a TVEMF sleeve being introduced to a rotatable culture container; and Figure 8 is a cross sectional elevated side view of a TVEMF sleeve removably adjacent to and encompassing a rotatable culture container.
DETAILED DESCRIPTION OF THE DRAWINGS
In the drawings, Figures 1-6 illustrate preferred embodiments of a TVEMF
sleeve that may supply a TVEMF to cells inside a culture container. Figures 7and 8 illustrate a preferred embodiment of a TVEMF sleeve 10 that is rotatable and that may supply a TVEMF to cells inside a rotatable culture container.
Figure 1 is an elevated side view of a preferred embodiment of the TVEMF
sleeve 10 comprising an electrically conductive coil 5 and a TVEMF source 9 operatively connected to the electrically conductive coil 5. The phrase "operatively connected," and similar words and phrases, is intended to mean that the TVEMF source can be connected to the electrically conductive coil in a manner such that when in operation, the TVEMF
source can impart a TVEMF to the electrically conductive coil through a conductive connection, preferably with at least one electrically conductive wire. The TVEMF source may preferably be affixed to the electrically conductive coil, and may preferably be integral with the electrically conductive coil. In the present invention, the electrically conductive coil may be any material that conducts electricity. The electrically conductive coil may preferably be, but is not limited to, the following conductive materials; silver, gold, copper, aluminum, iron, lead, titanium, uranium, a ferromagnetic metal,.. and zinc, or a combination thereof. The electrically conductive coil may also preferably comprise salt water. The electrically conductive coil of the present invention may preferably be substantially rigid. "Substantially rigid," is intended throughout this invention to mean that the electrically conductive coil can maintain a shape without the need for support.
The electrically conductive coil may be any shape, preferably substantially cylindrical preferably with a substantially elliptical cross-section, more preferably with a substantially oval cross-section, and most preferably with a substantially circular cross-section. The electrically conductive coil may also preferably be a solenoid, a tightly wound electrically conductive coil. Furthermore, the electrically conductive coil may preferably be wrapped in electric insulators comprising, but not limited to, rubber, plastic, silicones, glass, and ceramic. Preferably the electric insulator provides rigidity to the electrically conductive coil.
In the preferred embod'unent illustrated in Figure 1, the electrically conductive coil 5 is wrapped around the exterior portion of a coil support 3. The coil support of the present invention may preferably be made of a non-conductive non-magnetic material, more preferably non-metallic, and most preferably plastic for example polyethylene.
Furthermore, the coil support may preferably comprise a conductive material, preferably iron. Not to be bound by the theory, an iron core is thought to enhance the magnetic field of a solenoid. The exterior portion of the coil support has a shape that is compatible with the electrically conductive coil. By "compatible" it is meant that the coil support lends support to the electrically conductive coil and preferably dictates and maintains the shape of the electrically conductive coil. The electrically conductive coil may be wound around the exterior portion of the coil support supporting a shape of the electrically conductive coil, preferably having a substantially oval cross-section, more preferably a substantially elliptical cross-section, and most preferably a substantially circular cross-section. In this preferred embodiment of the TVEMF sleeve 10, the electrically conductive coil 5 and the coil support 3 are disposed on, preferably removably, a base 11 that provides stationary support thereto.
Figure 2 is an elevated front view of a preferred embodiment of the TVEMF
sleeve 10, also illustrated in Figure 1, depicting the electrically conductive coil 5 wrapped around the exterior portion of the coil support 3 and wherein the electrically conductive coil 5 and the coil support 3 are disposed on, preferably removably, a base 11.
Figure 3 is an elevated front view of a preferred embodiment of the TVEMF
sleeve 10, also illustrated in Figures 1 and 2, further having a culture container 1 in the interior portion of the coil support 3 wound with the electrically conductive coil 5. In the present invention, the electrically conductive coil, and the coil support wlien present, has an interior portion that defines a space in which a culture container is removably received. "Removably received" and similar terms refers to a characteristic of a space wherein a culture container is introduced thereto and rerrioved there from as desired. For example, a culture container is removably received by the space in the interior portion of the electrically conductive coil or the coil support when present. The space can accommodate a culture container so that the culture container may be removed and /or dissembled from the space as desired. The space in the interior portion of the coil support may be any shape and preferably has a side on which at least one culture container can be removably disposed.
The culture container of the present invention may be a culture container which can be encompassed by, and contained within, the interior portion of the electrically conductive coil, and coil support where applicable, including, but not limited to, a bioreactor culture container, a rotatable bioreactor culture container, a flask, a plate, a petri dish, and/or a disposable culture container. By "encompasses" it is meant that the electrically conductive coil, and the coil support where applicable, surrounds the culture container. Furkhermore, the culture container may be a single culture container or more than one culture container. The culture container of the present invention is also capable of supporting and maintaining a cell culture, preferrably the expansion of a cell, cell aggregate, tissue, or tissue-like structure.
Figure 4 is an elevated side view of another preferred embodiment of the TVEMF
sleeve 10 of the present invention having an electrically conductive coil 105 and an operatively connected TVEMW source 109. The electrically conductive coil 105 is wrapped in a substantially cylindrical shape, preferably witli a substantially oval cross-section, and more preferably with a substantially circular cross-section, and inost preferably with a substantially elliptical cross-section. The electrically conductive coil 105 in this preferred embodiment is substantially rigid and is preferably insulated. The electric insulator may preferably provide rigidity to the shape of the electrically conductive coil. The electrically conductive coil of the TVEMF sleeve 10 has a cross-section that is large enough to removably receive and encompass a culture container.
Furthermore, the electrically conductive coil 105 is removably adjacent to the culture container 101. By "removably adjacent to," and similar terms, it is meant that the electrically conductive coil can be next to, near to, or touching the culture container, and so that it can be easily removed from, or disassembled from, the culture container.
Figure 5 illustrates an elevated side view of yet another preferred embodiment of the TVEMF sleeve 10 having an electrically conductive coi1205 and a TVEMF source operatively connected to the electrically conductive coi1205. In this preferred embodiment of the TVEMF sleeve 10, the electrically conductive coil 205 is wrapped around the exterior portion of a coil support 203. Figure 5 further illustrates the TVEMF sleeve 10 being introduced to a culture container 201, for instance a petri dish. In this preferred embodiment, the culture container 201 is placed on a riser 213, which is in turn disposed on a base 211, preferably removably disposed on, so that the riser 213 may be removed from the base 211 and may be cleaned, sterilized, and/or otherwise maintained as needed. The electrically conductive coil 205 is disposed on, preferably removably, the base 211. To introduce the culture container 201 to the TVEMF sleeve 10, the culture container 201 is placed on the riser 213, and the electrically conductive coil 205 and coil support 203 are then placed over the culture container 201. The term "introducing," and similar terms, is intended to refer to the removable reception and accommodation of a culture container in the interior portion of the electrically conductive coil, or the coil support where applicable. Manipulating a TVEMF sleeve so that it encompasses, and is removably adjacent to, a culture container is considered introducing a TVEMF
sleeve to a culture container. Furthermore, manipulating a culture container so that it is encompassed by a TVEMF sleeve is also considered introducing a culture container to a TVEMF
sleeve. In the present invention, in use, once introduced, the electrically conductive coil of the TVEMF sleeve is removably adjacent to the culture container and at the same time encompasses the culture container.
Figure 6 is an elevated side view of the preferred embodiment of the TVEMF
sleeve 10 illustrated in Figure 5. The TVEMF sleeve 10 comprises an electrically conductive coil 205 and an operatively connected TVEMF source 209. In Figure 6, as in Figure 5, the electrically conductive coil 205 is wrapped around the exterior portion of a coil support 203. The coil support 203 is removably adjacent to, and encompasses, a culture container and the coil support 203, wound with the electrically conductive coil 205, is removably disposed on a base 211.
Figure 7 is a cross sectional elevated side view of another preferred einbodiment of a TVEMF sleeve 10 and its introduction to a rotatable culture container 301. The TVEMF sleeve 10 of the preferred embodiment in Figure 6 comprises an electrically conductive coil 305 and a TVEMF source 309 operatively connected to the electrically conductive coil 305. In this preferred embodiment, the electrically conductive coil 305 is wrapped around the exterior portion of a coil support 303 and the TVEMF sleeve 10 is rotatable. At a first end a first conductive wire 325 and a second conductive wire 326 are connected to the TVEMF source 309. At a second end the wires 325, 326 are connected to at least one ring to facilitate the rotation of the electrically conductive coil 305, preferably a first ring 321 and a second ring 322 respectively. Also in Figure 7 is depicted a motor housing 312 supported by a base 311. A motor 313 is affixed inside the motor housing 312 and connected by a first wire 314 and a second wire 315 to a control box 316 that houses a control device therein whereby the speed of the motor 313 can be incrementally controlled by turning the control knob 317. Extending from the motor housing 312 is a motor shaft 318. A rotatable mounting 328 removably receives a rotatable culture container holder 329, preferably disposable, that removeably receives a rotatable culture container 301 which is removably affixed within the rotatable culture container holder 329, preferably by a screw 331.
In Figure 7, preferably the TVEMF sleeve 10 and the rotatable culture container 301 are removably mounted to the rotatable mounting 328. The rotatable mounting 328 is received by the motor shaft 318. When the control knob 317 is turned on, the rotatable culture container 301 along with the coil support 303, wound by the electrically conductive coil 305, may preferably be rotated siinultaneously. Furthennore, in operation the TVEIvLF sleeve 10 remains removably adjacent to, and encompasses, the rotatable culture container 301, while at the same time, supplying a TVEMF to the cells in the rotatable culture container 301. The rotatable culture container may preferably be disposable wllerein it can be discarded and a new one used in later cell cultures. The rotatable culture container may also preferably be sterilized, for instance in an autoclave, after each use and re-used for later cell cultures. A disposable culture container could be manufactured and packaged in a sterile environment thereby enabling it to be used by the medical or research professional much the same as other disposable medical devices are used.
Figure 8 illustrates a cross-sectional elevated side view of the TVEMF sleeve in the preferred embodiment of Figure 7 removably adjacent to, and encompassing, a rotatable culture container 301.
In operation, a culture container is introduced to a TVEMF sleeve. The TVEMF
source of the TVEMF sleeve is turned on and a TVEMF is delivered through the electrically conductive coil into a culture container encompassed by, and removably adjacent to, the TVEMF sleeve. In use, the TVEMF is distributed throughout the culture container, and therefore, to the cells contained therein, preferably in a nearly uniform distribution. In the present invention, the term "cells," or any other term similar term, is intended to include, but is not limited to single cells, cells attached to cell attachment substrates, cell aggregates, tissues, and tissue-like structures. The term "expansion" is intended to include growth in the size of tissue(s), tissue-like structures, and/or cell aggregates, and/or the growth in the number of cells in a culture container.
Because the present invention provides a method for supplying a TVEMF to cells in a culture container for expansion of the same, a TVEMF sleeve is so sized and configured to removably receive the culture container so that a TVEMF can be supplied to the cells in the culture container, preferably in a nearly uniform distribution. Since the TVEMF sleeve of the present invention is removably adjacent to the culture container, the TVEMF sleeve may be reused for sequential cell cultures. Also, because the TVEMF
sleeve is removably adjacent to the culture container, a single TVEMF sleeve can accommodate different types, shapes, and sizes of culture containers.
The size of the electrically conductive coil and the number of times it is wound are such that when a TVEMF is supplied to the electrically conductive coil, a TVEMF is generated in the culture container for the expansion of cells therein. The TVEMF sleeve may generate a TVEMF preferably of from about 0.05 gauss to about 6 gauss, more preferably of from about 0.05 gauss to about 0.5 gauss, and most preferably about 0.5 gauss. The TVEMF is preferably in a delta wave, more preferably in a differentiated square wave, and most preferably in a square wave (following a Fourier curve).
Preferably, the pulsed square wave has a frequency of about 2 to about 25 cycles/second, more preferably about 5 to about 20 cycles/second, and for example about 10 cycles/second, and the electrically conductive coil has an RMS value of about 1 to about 1000 mA, preferably about 1 to about 10 mA, for example 6 mA. However, these parameters are not meant to be limiting to the TVEMF of the present invention, and as such, may vary based on other aspects of this invention. TVEMF may be measured by standard equipment, for instance an EN331 Cell Sensor Gauss Meter.
It is fiirther contemplated that the TVEMF sleeve may be equipped with a temperature control device. The temperature control device, preferably be an automated sensor, detects the temperature in the interior portion of the electrically conductive coil, and coil support where applicable. In use, if the temperature control device detects a temperature reading in the interior portion of the electrically conductive coil, and coil support when present, that is higher than desired, the temperature control device may alert the user of the temperature change, preferably with an alarm. Not to be bound by the theory, but when there is a high electrical resistance, heat is generated.
Thus, the space wherein a culture container is-removably received may become hot depending upon the diameter of the electrically conductive coil and the amount of electricity supplied thereto.
EXPANSION OF CELLS AND METHOD OF USING THE SAME
FIELD OF THE INVENTION
The present invention relates generally to a sleeve for the expansion of cells, and more particularly to a time va.rying electroinagnetic force sleeve that removably recieves a culture container. The present invention also relates to a time varying electromagnetic force sleeve that, when in use, supplies a time varying electromagnetic force to cells in a culture container for cell expansion therein.
BACKGROUND OF THE INVENTION
For many years there have existed methods for culturing cells. Some methods involve culturing cells in two-dimensional cultures utilizing such culture chambers as flasks and petri dishes, and others involve culturing cells in three-dimensional cultures utilizing such culture chambers as bioreactors. Methods of optimally culturing cells include adding molecules to cells in a culture such as growth factors, hormones, and others that, for instance, up or down regulate expansion of cells. Some methods are optimized for culturing individual cells and others are optimized for tissue culture. In addition, in an effort to produce more cells or larger tissue constructs over time, cell cultures are performed under various conditions.
United States Patent Number 6,673,597, Wolf et al., disclose the use of a time varying electromagnetic force ("TVEMF") to grow cells in culture. Wolf et al., disclose the use of electrodes to supply an electromagnetic force to a cell culture for growth of the same. In those instances where a conductive coil, rather than electrodes, is used for the delivery of a TVEMF to a cell culture, the coil is integral with a culture chamber and affixed thereto. Since the cells expanded in a culture may be introduced into the human body for tissue regeneration or treatment of human maladies, the culture chamber should be sterile. If the source of TVEMF to be delivered to cells in a culture chamber is integral with the culture chamber then either the culture chamber should be sterilized without disturbing the source of the TVEMF, a process which is cumbersome and time consuming, and if the container is to be discarded after each use, then the integral TVEMF coil has to be discarded therewith. Either way, an integral TVEMF coil is an expensive approach to cell expansion.
Consequently, it would be highly desirable to have a TVEMF sleeve comprising an electrically conductive coil having an interior portion wherein the interior portion defines a space wherein a culture container is removably received. It is also highly desirable to have a TVEMF sleeve comprising a coil support with an interior portion and an exterior portion, an electrically conductive coil wrapped around the exterior portion of the coil support; and a TVEMF source, and wherein the interior portion of the coil support defines a space wherein a culture container is removably received. It is fu.rther highly desirable to have a TVEMF sleeve that, when in use, is introduced to a culture container containing cells and supplies a TVEMF to the cells in a culture container for cell expansion. The present invention overcomes the problems associated with past and current cell culture methods, and presents advantages not before seen.
SUMMARY OF THE INVENTION
The present invention relates to a TVEMF sleeve comprising an electrically conductive coil having an interior portion and an exterior portion wherein the interior portion defines a space wherein a culture container is removably received, and a TVEMF
source operatively connected to the electrically conductive coil. Preferably the electrically conductive coil is substantially rigid.
The present invention also relates to a TVEMF sleeve comprising a coil support with an interior portion and an exterior portion wherein the interior portion defines a space in which a culture container is removably received, an electrically conductive coil wrapped around the exterior portion of the coil support, and a TVEMF source operatively connected to the electrically conductive coil.
The present invention also relates to a method of cell expansion comprising the steps of providing a time varying electromagnetic force sleeve comprising an electrically conductive coil, having an interior portion and exterior portion, operatively connected to a time varying electromagnetic force source; introducing a culture container containing cells to the interior portion of the electrically conductive coil of the time varying electromagnetic force sleeve; and supplying a time varying electromagnetic force to the cells in the culture container through the time varying electromagnetic force sleeve for cell expansion. The TVEMF sleeve may preferably deliver a TVEMF in the form of a pulsed square wave (following a Fourier curve) to the cells in the culture container, and the TVEMF may preferably be of from about 0.05 gauss to about 6 gauss.
Other aspects, features, and advantages of the present invention will be apparent from the following description of the preferred embodiments of the invention given for the purpose of disclosure. This invention may be more fully described by the preferred embodiment(s) as hereinafter described, but is not intended to be limited thereto.
BRIEF DESCRIPTION OF THE D.RAWINGS
In the drawings, Figure 1 is an elevated side view of a TVEMF sleeve;
Figure 2 is an elevated front view of a TVEMF sleeve;
Figure 3 is an elevated front view of a TVEMF sleeve removably adjacent to and encompassing a culture container;
Figure 4 is an elevated side view of a TVEMF sleeve;
Figure 5 is an elevated side view of a TVEMF sleeve being introduced to a culture container;
Figure 6 is an elevated side view of a TVEMF sleeve;
Figure 7 is a cross sectional elevated side view of a TVEMF sleeve being introduced to a rotatable culture container; and Figure 8 is a cross sectional elevated side view of a TVEMF sleeve removably adjacent to and encompassing a rotatable culture container.
DETAILED DESCRIPTION OF THE DRAWINGS
In the drawings, Figures 1-6 illustrate preferred embodiments of a TVEMF
sleeve that may supply a TVEMF to cells inside a culture container. Figures 7and 8 illustrate a preferred embodiment of a TVEMF sleeve 10 that is rotatable and that may supply a TVEMF to cells inside a rotatable culture container.
Figure 1 is an elevated side view of a preferred embodiment of the TVEMF
sleeve 10 comprising an electrically conductive coil 5 and a TVEMF source 9 operatively connected to the electrically conductive coil 5. The phrase "operatively connected," and similar words and phrases, is intended to mean that the TVEMF source can be connected to the electrically conductive coil in a manner such that when in operation, the TVEMF
source can impart a TVEMF to the electrically conductive coil through a conductive connection, preferably with at least one electrically conductive wire. The TVEMF source may preferably be affixed to the electrically conductive coil, and may preferably be integral with the electrically conductive coil. In the present invention, the electrically conductive coil may be any material that conducts electricity. The electrically conductive coil may preferably be, but is not limited to, the following conductive materials; silver, gold, copper, aluminum, iron, lead, titanium, uranium, a ferromagnetic metal,.. and zinc, or a combination thereof. The electrically conductive coil may also preferably comprise salt water. The electrically conductive coil of the present invention may preferably be substantially rigid. "Substantially rigid," is intended throughout this invention to mean that the electrically conductive coil can maintain a shape without the need for support.
The electrically conductive coil may be any shape, preferably substantially cylindrical preferably with a substantially elliptical cross-section, more preferably with a substantially oval cross-section, and most preferably with a substantially circular cross-section. The electrically conductive coil may also preferably be a solenoid, a tightly wound electrically conductive coil. Furthermore, the electrically conductive coil may preferably be wrapped in electric insulators comprising, but not limited to, rubber, plastic, silicones, glass, and ceramic. Preferably the electric insulator provides rigidity to the electrically conductive coil.
In the preferred embod'unent illustrated in Figure 1, the electrically conductive coil 5 is wrapped around the exterior portion of a coil support 3. The coil support of the present invention may preferably be made of a non-conductive non-magnetic material, more preferably non-metallic, and most preferably plastic for example polyethylene.
Furthermore, the coil support may preferably comprise a conductive material, preferably iron. Not to be bound by the theory, an iron core is thought to enhance the magnetic field of a solenoid. The exterior portion of the coil support has a shape that is compatible with the electrically conductive coil. By "compatible" it is meant that the coil support lends support to the electrically conductive coil and preferably dictates and maintains the shape of the electrically conductive coil. The electrically conductive coil may be wound around the exterior portion of the coil support supporting a shape of the electrically conductive coil, preferably having a substantially oval cross-section, more preferably a substantially elliptical cross-section, and most preferably a substantially circular cross-section. In this preferred embodiment of the TVEMF sleeve 10, the electrically conductive coil 5 and the coil support 3 are disposed on, preferably removably, a base 11 that provides stationary support thereto.
Figure 2 is an elevated front view of a preferred embodiment of the TVEMF
sleeve 10, also illustrated in Figure 1, depicting the electrically conductive coil 5 wrapped around the exterior portion of the coil support 3 and wherein the electrically conductive coil 5 and the coil support 3 are disposed on, preferably removably, a base 11.
Figure 3 is an elevated front view of a preferred embodiment of the TVEMF
sleeve 10, also illustrated in Figures 1 and 2, further having a culture container 1 in the interior portion of the coil support 3 wound with the electrically conductive coil 5. In the present invention, the electrically conductive coil, and the coil support wlien present, has an interior portion that defines a space in which a culture container is removably received. "Removably received" and similar terms refers to a characteristic of a space wherein a culture container is introduced thereto and rerrioved there from as desired. For example, a culture container is removably received by the space in the interior portion of the electrically conductive coil or the coil support when present. The space can accommodate a culture container so that the culture container may be removed and /or dissembled from the space as desired. The space in the interior portion of the coil support may be any shape and preferably has a side on which at least one culture container can be removably disposed.
The culture container of the present invention may be a culture container which can be encompassed by, and contained within, the interior portion of the electrically conductive coil, and coil support where applicable, including, but not limited to, a bioreactor culture container, a rotatable bioreactor culture container, a flask, a plate, a petri dish, and/or a disposable culture container. By "encompasses" it is meant that the electrically conductive coil, and the coil support where applicable, surrounds the culture container. Furkhermore, the culture container may be a single culture container or more than one culture container. The culture container of the present invention is also capable of supporting and maintaining a cell culture, preferrably the expansion of a cell, cell aggregate, tissue, or tissue-like structure.
Figure 4 is an elevated side view of another preferred embodiment of the TVEMF
sleeve 10 of the present invention having an electrically conductive coil 105 and an operatively connected TVEMW source 109. The electrically conductive coil 105 is wrapped in a substantially cylindrical shape, preferably witli a substantially oval cross-section, and more preferably with a substantially circular cross-section, and inost preferably with a substantially elliptical cross-section. The electrically conductive coil 105 in this preferred embodiment is substantially rigid and is preferably insulated. The electric insulator may preferably provide rigidity to the shape of the electrically conductive coil. The electrically conductive coil of the TVEMF sleeve 10 has a cross-section that is large enough to removably receive and encompass a culture container.
Furthermore, the electrically conductive coil 105 is removably adjacent to the culture container 101. By "removably adjacent to," and similar terms, it is meant that the electrically conductive coil can be next to, near to, or touching the culture container, and so that it can be easily removed from, or disassembled from, the culture container.
Figure 5 illustrates an elevated side view of yet another preferred embodiment of the TVEMF sleeve 10 having an electrically conductive coi1205 and a TVEMF source operatively connected to the electrically conductive coi1205. In this preferred embodiment of the TVEMF sleeve 10, the electrically conductive coil 205 is wrapped around the exterior portion of a coil support 203. Figure 5 further illustrates the TVEMF sleeve 10 being introduced to a culture container 201, for instance a petri dish. In this preferred embodiment, the culture container 201 is placed on a riser 213, which is in turn disposed on a base 211, preferably removably disposed on, so that the riser 213 may be removed from the base 211 and may be cleaned, sterilized, and/or otherwise maintained as needed. The electrically conductive coil 205 is disposed on, preferably removably, the base 211. To introduce the culture container 201 to the TVEMF sleeve 10, the culture container 201 is placed on the riser 213, and the electrically conductive coil 205 and coil support 203 are then placed over the culture container 201. The term "introducing," and similar terms, is intended to refer to the removable reception and accommodation of a culture container in the interior portion of the electrically conductive coil, or the coil support where applicable. Manipulating a TVEMF sleeve so that it encompasses, and is removably adjacent to, a culture container is considered introducing a TVEMF
sleeve to a culture container. Furthermore, manipulating a culture container so that it is encompassed by a TVEMF sleeve is also considered introducing a culture container to a TVEMF
sleeve. In the present invention, in use, once introduced, the electrically conductive coil of the TVEMF sleeve is removably adjacent to the culture container and at the same time encompasses the culture container.
Figure 6 is an elevated side view of the preferred embodiment of the TVEMF
sleeve 10 illustrated in Figure 5. The TVEMF sleeve 10 comprises an electrically conductive coil 205 and an operatively connected TVEMF source 209. In Figure 6, as in Figure 5, the electrically conductive coil 205 is wrapped around the exterior portion of a coil support 203. The coil support 203 is removably adjacent to, and encompasses, a culture container and the coil support 203, wound with the electrically conductive coil 205, is removably disposed on a base 211.
Figure 7 is a cross sectional elevated side view of another preferred einbodiment of a TVEMF sleeve 10 and its introduction to a rotatable culture container 301. The TVEMF sleeve 10 of the preferred embodiment in Figure 6 comprises an electrically conductive coil 305 and a TVEMF source 309 operatively connected to the electrically conductive coil 305. In this preferred embodiment, the electrically conductive coil 305 is wrapped around the exterior portion of a coil support 303 and the TVEMF sleeve 10 is rotatable. At a first end a first conductive wire 325 and a second conductive wire 326 are connected to the TVEMF source 309. At a second end the wires 325, 326 are connected to at least one ring to facilitate the rotation of the electrically conductive coil 305, preferably a first ring 321 and a second ring 322 respectively. Also in Figure 7 is depicted a motor housing 312 supported by a base 311. A motor 313 is affixed inside the motor housing 312 and connected by a first wire 314 and a second wire 315 to a control box 316 that houses a control device therein whereby the speed of the motor 313 can be incrementally controlled by turning the control knob 317. Extending from the motor housing 312 is a motor shaft 318. A rotatable mounting 328 removably receives a rotatable culture container holder 329, preferably disposable, that removeably receives a rotatable culture container 301 which is removably affixed within the rotatable culture container holder 329, preferably by a screw 331.
In Figure 7, preferably the TVEMF sleeve 10 and the rotatable culture container 301 are removably mounted to the rotatable mounting 328. The rotatable mounting 328 is received by the motor shaft 318. When the control knob 317 is turned on, the rotatable culture container 301 along with the coil support 303, wound by the electrically conductive coil 305, may preferably be rotated siinultaneously. Furthennore, in operation the TVEIvLF sleeve 10 remains removably adjacent to, and encompasses, the rotatable culture container 301, while at the same time, supplying a TVEMF to the cells in the rotatable culture container 301. The rotatable culture container may preferably be disposable wllerein it can be discarded and a new one used in later cell cultures. The rotatable culture container may also preferably be sterilized, for instance in an autoclave, after each use and re-used for later cell cultures. A disposable culture container could be manufactured and packaged in a sterile environment thereby enabling it to be used by the medical or research professional much the same as other disposable medical devices are used.
Figure 8 illustrates a cross-sectional elevated side view of the TVEMF sleeve in the preferred embodiment of Figure 7 removably adjacent to, and encompassing, a rotatable culture container 301.
In operation, a culture container is introduced to a TVEMF sleeve. The TVEMF
source of the TVEMF sleeve is turned on and a TVEMF is delivered through the electrically conductive coil into a culture container encompassed by, and removably adjacent to, the TVEMF sleeve. In use, the TVEMF is distributed throughout the culture container, and therefore, to the cells contained therein, preferably in a nearly uniform distribution. In the present invention, the term "cells," or any other term similar term, is intended to include, but is not limited to single cells, cells attached to cell attachment substrates, cell aggregates, tissues, and tissue-like structures. The term "expansion" is intended to include growth in the size of tissue(s), tissue-like structures, and/or cell aggregates, and/or the growth in the number of cells in a culture container.
Because the present invention provides a method for supplying a TVEMF to cells in a culture container for expansion of the same, a TVEMF sleeve is so sized and configured to removably receive the culture container so that a TVEMF can be supplied to the cells in the culture container, preferably in a nearly uniform distribution. Since the TVEMF sleeve of the present invention is removably adjacent to the culture container, the TVEMF sleeve may be reused for sequential cell cultures. Also, because the TVEMF
sleeve is removably adjacent to the culture container, a single TVEMF sleeve can accommodate different types, shapes, and sizes of culture containers.
The size of the electrically conductive coil and the number of times it is wound are such that when a TVEMF is supplied to the electrically conductive coil, a TVEMF is generated in the culture container for the expansion of cells therein. The TVEMF sleeve may generate a TVEMF preferably of from about 0.05 gauss to about 6 gauss, more preferably of from about 0.05 gauss to about 0.5 gauss, and most preferably about 0.5 gauss. The TVEMF is preferably in a delta wave, more preferably in a differentiated square wave, and most preferably in a square wave (following a Fourier curve).
Preferably, the pulsed square wave has a frequency of about 2 to about 25 cycles/second, more preferably about 5 to about 20 cycles/second, and for example about 10 cycles/second, and the electrically conductive coil has an RMS value of about 1 to about 1000 mA, preferably about 1 to about 10 mA, for example 6 mA. However, these parameters are not meant to be limiting to the TVEMF of the present invention, and as such, may vary based on other aspects of this invention. TVEMF may be measured by standard equipment, for instance an EN331 Cell Sensor Gauss Meter.
It is fiirther contemplated that the TVEMF sleeve may be equipped with a temperature control device. The temperature control device, preferably be an automated sensor, detects the temperature in the interior portion of the electrically conductive coil, and coil support where applicable. In use, if the temperature control device detects a temperature reading in the interior portion of the electrically conductive coil, and coil support when present, that is higher than desired, the temperature control device may alert the user of the temperature change, preferably with an alarm. Not to be bound by the theory, but when there is a high electrical resistance, heat is generated.
Thus, the space wherein a culture container is-removably received may become hot depending upon the diameter of the electrically conductive coil and the amount of electricity supplied thereto.
As various changes could be made in TVEMF sleeves, as are contemplated in the present invention, without departing from the scope of the invention, it is intended that all matter contained herein be interpreted as illustrative and not limiting.
OPERATIVE METHOD
Peripheral blood cells PBCs were collected from donors and a culture mix was made with the whole blood that was collected wherein the cells (0.75 x 106 cells/ml) were suspended in Iscove's modified Dulbecco's medium (IMDM) (GIBCO, Grand Island, NY) supplemented with 5% human albumin (HA), 100 ng/ml recombinant human G-CSF (Amgen Inc., Thousand Oaks, CA), and 100 ng/inl recombinant human stem cell factor (SCF) (Amgen). 10 ppm of D-Penicillamine [D(-)-2-Amino-3-mercapto-3-methylbutanoic acid] (Sigma-Aldrich), a copper chelating agent, dissolved in DMSO, was introduced into the cell mixture. The purpose of the copper chelating ageiit is, not to be bound by theory, to reduce the amount of copper in the peripheral blood prior to TVEMF- expansion. Not to be bound by theory, it is believed that the decrease in amount of available copper may enhance cell expansion.
Example 1- Cell expansion in a rotatable bioreactor A first sample of the culture mix, prepared as above, was placed into a rotatable bioreactor having a 75 ml culture chamber. 0.75 x 106 cells/ml of the culture mix, was placed in the 75 ml culture chamber, for a total culture mixture volume of 75 ml. A time varying electromagnetic force of about 0.5 gauss in a pulsed square wave was supplied to the cells in the culture container of the rotatable bioreactor through the TVEMF sleeve encompassing and removably adajacent to the rotatable bioreactor, for example in Figures 7 and S. A second sample was placed in a rotatable bioreactor without any TVEMF applied thereto.
Other than the TVEMF condition, all other conditions were identical as between the first and second sample.
The culture containers were rotated at a speed of about 10 RPM. The cultures were grown at 37 C and in 5% COa.
The cells of the first and second sample were allowed to expand for seven days and after the seventh day of expansion the cells were washed with PBS
(Phosphate Buffer Solution) and counted by conventional counting techniques, for example by using a Coulter cell counter. By visual determination, it was found that the first culture mix that was exposed to a TVEMF via the TVEMF sleeve had more than five times the growth or expansion of the sample that was not exposed to a TVEMF.
Example 2- Cells expansion in a uetri dish A first sample of the culture mix, prepared as above, was placed into a petri dish and a TVEMF of about 0.5 gauss, in a pulsed square wave, was supplied to the TVEMF
sleeve that encompassed, and was adjacent to, the petri dish, for example in Figures 5 and 6. A second sample was placed in a petri dish without any TVEMF applied thereto. Other than the TVEMF
condition, all other conditions were identical asbetween the first and second sample. The cultures were grown in an incubator at 37 C and in 5% CO2.
The cells of the first and second sample were allowed to expand for seven days and after the seventh day of expansion the cells were washed with PBS
(Phosphate Buffer Solution) and counted by conventional counting techniques, for example by using a Coulter cell counter. By visual determination it was found that the culture mix that was exposed to a TVEMF via the TVEMF sleeve had more than two times the growth or expansion of the sample that was not exposed to a TVEMF.
OPERATIVE METHOD
Peripheral blood cells PBCs were collected from donors and a culture mix was made with the whole blood that was collected wherein the cells (0.75 x 106 cells/ml) were suspended in Iscove's modified Dulbecco's medium (IMDM) (GIBCO, Grand Island, NY) supplemented with 5% human albumin (HA), 100 ng/ml recombinant human G-CSF (Amgen Inc., Thousand Oaks, CA), and 100 ng/inl recombinant human stem cell factor (SCF) (Amgen). 10 ppm of D-Penicillamine [D(-)-2-Amino-3-mercapto-3-methylbutanoic acid] (Sigma-Aldrich), a copper chelating agent, dissolved in DMSO, was introduced into the cell mixture. The purpose of the copper chelating ageiit is, not to be bound by theory, to reduce the amount of copper in the peripheral blood prior to TVEMF- expansion. Not to be bound by theory, it is believed that the decrease in amount of available copper may enhance cell expansion.
Example 1- Cell expansion in a rotatable bioreactor A first sample of the culture mix, prepared as above, was placed into a rotatable bioreactor having a 75 ml culture chamber. 0.75 x 106 cells/ml of the culture mix, was placed in the 75 ml culture chamber, for a total culture mixture volume of 75 ml. A time varying electromagnetic force of about 0.5 gauss in a pulsed square wave was supplied to the cells in the culture container of the rotatable bioreactor through the TVEMF sleeve encompassing and removably adajacent to the rotatable bioreactor, for example in Figures 7 and S. A second sample was placed in a rotatable bioreactor without any TVEMF applied thereto.
Other than the TVEMF condition, all other conditions were identical as between the first and second sample.
The culture containers were rotated at a speed of about 10 RPM. The cultures were grown at 37 C and in 5% COa.
The cells of the first and second sample were allowed to expand for seven days and after the seventh day of expansion the cells were washed with PBS
(Phosphate Buffer Solution) and counted by conventional counting techniques, for example by using a Coulter cell counter. By visual determination, it was found that the first culture mix that was exposed to a TVEMF via the TVEMF sleeve had more than five times the growth or expansion of the sample that was not exposed to a TVEMF.
Example 2- Cells expansion in a uetri dish A first sample of the culture mix, prepared as above, was placed into a petri dish and a TVEMF of about 0.5 gauss, in a pulsed square wave, was supplied to the TVEMF
sleeve that encompassed, and was adjacent to, the petri dish, for example in Figures 5 and 6. A second sample was placed in a petri dish without any TVEMF applied thereto. Other than the TVEMF
condition, all other conditions were identical asbetween the first and second sample. The cultures were grown in an incubator at 37 C and in 5% CO2.
The cells of the first and second sample were allowed to expand for seven days and after the seventh day of expansion the cells were washed with PBS
(Phosphate Buffer Solution) and counted by conventional counting techniques, for example by using a Coulter cell counter. By visual determination it was found that the culture mix that was exposed to a TVEMF via the TVEMF sleeve had more than two times the growth or expansion of the sample that was not exposed to a TVEMF.
Claims (46)
1. A time varying electromagnetic force sleeve comprising:
-an electrically conductive coil having an interior portion and an exterior portion wherein the interior portion defines a space in which a rotatable bioreactor is removably received; and -a time varying electromagnetic force source operatively connected to the electrically conductive coil.
-an electrically conductive coil having an interior portion and an exterior portion wherein the interior portion defines a space in which a rotatable bioreactor is removably received; and -a time varying electromagnetic force source operatively connected to the electrically conductive coil.
2. A time varying electromagnetic force sleeve as in claim 1, wherein the electrically conductive coil is substantially rigid.
3. A time varying electromagnetic force sleeve as in claim 1, wherein the time varying electromagnetic force sleeve is rotatable about an axis.
4. A time varying electromagnetic force sleeve as in claim 3, wherein the axis is substantially horizontal.
5. A time varying electromagnetic force sleeve as in claim 3, wherein the axis is substantially vertical.
6. A time varying electromagnetic force sleeve as in claim 1, wherein the electrically conductive coil is a solenoid.
7. A time varying electromagnetic force sleeve as in claim 1, wherein the electrically conductive coil is substantially cylindrical.
8. A time varying electromagnetic force sleeve as in claim 1, wherein the electrically conductive coil has a substantially circular cross-section.
9. A time varying electromagnetic force sleeve as in claim 1, wherein the electrically conductive coil has a substantially oval cross-section.
10. A time varying electromagnetic force sleeve as in claim 1, wherein the electrically conductive coil has a substantially elliptical cross-section.
11. A time varying electromagnetic force sleeve as in claim 1, wherein the culture container is rotatable about an axis.
12. A time varying electromagnetic force sleeve as in claim 1, wherein the electrically conductive coil is insulated.
13. A time varying electromagnetic force sleeve as in claim 1, further comprising a coil support having an interior portion wherein the coil support is polyethylene, wherein the coil support is located in the interior portion of the electrically conductive coil, and wherein the interior portion of the coil support removably receives the culture container.
14. A time varying electromagnetic force sleeve comprising:
-a coil support having an interior portion and an exterior portion wherein the interior portion defines a space in which a rotatable bioreactor is removably received and wherein the coil support is polyethylene;
-an electrically conductive coil wound around the exterior portion of the coil support; and -a time varying electromagnetic force source operatively connected to the electrically conductive coil.
-a coil support having an interior portion and an exterior portion wherein the interior portion defines a space in which a rotatable bioreactor is removably received and wherein the coil support is polyethylene;
-an electrically conductive coil wound around the exterior portion of the coil support; and -a time varying electromagnetic force source operatively connected to the electrically conductive coil.
15. A time varying electromagnetic force sleeve as in claim 14, wherein the time varying electromagnetic force sleeve is rotatable about an axis.
16. A time varying electromagnetic force sleeve as in claim 15, wherein the axis is substantially horizontal.
17. A time varying electromagnetic force sleeve as in claim 15, wherein the axis is substantially vertical.
18. A time varying electromagnetic force sleeve as in claim 14, wherein the electrically conductive coil is a solenoid.
19. A time varying electromagnetic force sleeve as in claim 14, wherein the electrically conductive coil has a substantially circular cross-section.
20. A time varying electromagnetic force sleeve as in claim 14, wherein the electrically conductive coil has a substantially oval cross-section.
21. A time varying electromagnetic force sleeve as in claim 14, wherein the electrically conductive coil has an elliptical cross-section.
22. A time varying electromagnetic force sleeve as in claim 14, wherein the culture container is rotatable about an axis.
23. A time varying electromagnetic force sleeve as in claim 14, wherein the electrically conductive coil is insulated.
24. A time varying electromagnetic force sleeve as in claim 14, wherein the coil support is non-conductive.
25. A time varying electromagnetic force sleeve as in claim 14, wherein the coil support comprises a conductive material.
26. A time varying electromagnetic force sleeve as in claim 25, wherein the conductive material is iron.
27. A time varying electromagnetic force sleeve as in claim 14, further comprising a temperature control device for controlling the temperature in the culture container.
28. A method of cell expansion comprising the steps of:
-providing a time varying electromagnetic force sleeve having an interior portion and exterior portion;
-preparing a culture mix comprising peripheral blood cells;
-placing the culture mix into a culture container;
-introducing [[a]] the culture container containing peripheral blood cells to the interior portion of the time varying electromagnetic force sleeve; [[and]]
-supplying a time varying electromagnetic force to the cells in the culture container through the time varying electromagnetic force sleeve [[for]]; and -expanding the peripheral blood cells to a concentration in the culture container within an amount of time.
-providing a time varying electromagnetic force sleeve having an interior portion and exterior portion;
-preparing a culture mix comprising peripheral blood cells;
-placing the culture mix into a culture container;
-introducing [[a]] the culture container containing peripheral blood cells to the interior portion of the time varying electromagnetic force sleeve; [[and]]
-supplying a time varying electromagnetic force to the cells in the culture container through the time varying electromagnetic force sleeve [[for]]; and -expanding the peripheral blood cells to a concentration in the culture container within an amount of time.
29. The method of claim 28, wherein the interior portion of the time varying electromagnetic force sleeve further comprises a coil support having an interior portion and an exterior portion wherein an electrically conductive coil is wrapped around the exterior portion of the coil support and wherein the interior portion of the coil support defines a space that removably receives the culture container.
30. The method of claim 28, wherein the interior portion of the time varying electromagnetic force sleeve further comprises an electrically conductive coil having an interior portion and an exterior portion wherein the interior portion defines a space that removably receives the culture container.
31. The method of claim 28 or 29, wherein the time varying electromagnetic force is a square wave.
32. The method of claim 28 or 29, wherein the time varying electromagnetic force is of from about 0.05 gauss to about 6 gauss.
33. The method of claim 28 or 29, wherein the time varying electromagnetic force is of from about 0.05 gauss to about 0.5 gauss.
34. The method of claim 28 or 29, wherein the time varying electromagnetic force is about 0.5 gauss.
35. The method of claim 31, wherein the square wave has a frequency of from about 2 cycles/second to about 25 cycles/second.
36. The method of claim 31, wherein the square wave has a frequency of from about cycles/second to about 20 cycles/second.
37. The method of claim 31, wherein the square wave has a frequency of about cycles/second.
38. The method of claim 28 or 29, wherein the time varying electromagnetic force is a differentiated square wave.
39. The method of claim 28 or 29, wherein the time varying electromagnetic force is a delta wave.
40. The method of claim 28 or 29, wherein the peripheral blood cells that are expanded are peripheral blood adult stem cells.
41. The method of claim 40, wherein the expanding step is for at least seven days.
42. The method of claim 40, wherein the cells are expanded to at least two times the number that were placed into the culture container.
43. The method of claim 40, wherein the cells are expanded to at least five times the number that were placed into the culture container.
44. The method of claim 28 or 29, wherein the culture mix further comprises a copper chelating agent.
45. The method of claim 28 or 29, wherein the culture container is a rotatable bioreactor.
46. The method of claim 45, further comprising the step of rotating the rotatable bioreactor after the step of introducing the rotatable bioreactor to the sleeve.
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US11/169,613 | 2005-06-29 | ||
US11/169,386 | 2005-06-29 | ||
US11/169,386 US20080044890A1 (en) | 2004-06-30 | 2005-06-29 | Interchangable sleeve for enhancing proliferation of cells in a rotating bioreactor |
US11/169,613 US20060228795A1 (en) | 2004-06-30 | 2005-06-29 | Apparatus for enhancing proliferation of cells in a small-scale cell culturing container |
PCT/US2006/024241 WO2007005306A1 (en) | 2005-06-29 | 2006-06-22 | Time varying electromagnetic force sleeve for the expansion of cells and method of using the same |
Publications (1)
Publication Number | Publication Date |
---|---|
CA2613408A1 true CA2613408A1 (en) | 2007-01-11 |
Family
ID=37604790
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA002613408A Abandoned CA2613408A1 (en) | 2005-06-29 | 2006-06-22 | Time varying electromagnetic force sleeve for the expansion of cells and method of using the same |
Country Status (8)
Country | Link |
---|---|
EP (1) | EP1896566A1 (en) |
JP (1) | JP2008544754A (en) |
KR (1) | KR20080023755A (en) |
BR (1) | BRPI0613092A2 (en) |
CA (1) | CA2613408A1 (en) |
IL (1) | IL188443A0 (en) |
MX (1) | MX2008000272A (en) |
WO (1) | WO2007005306A1 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2023110063A1 (en) * | 2021-12-14 | 2023-06-22 | Azyro Sa | Apparatus for in-vitro influencing biological cellular material, and use thereof |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20130267003A1 (en) * | 2012-04-09 | 2013-10-10 | Thomas J. Goodwin | Methods for Culturing Cells in an Alternating Ionic Magnetic Resonance (AIMR) Multiple-Chambered Culture Apparatus |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH05219936A (en) * | 1992-02-14 | 1993-08-31 | Kobe Steel Ltd | Bioreactor |
JPH0670747A (en) * | 1992-08-24 | 1994-03-15 | Hitachi Ltd | Method for culturing cell and reacting enzyme |
JP3554220B2 (en) * | 1999-05-06 | 2004-08-18 | 独立行政法人 科学技術振興機構 | Fermentation method and fermentation apparatus |
US6485963B1 (en) * | 2000-06-02 | 2002-11-26 | The United States Of America As Represented By The Administrator Of The National Aeronautics And Space Administration | Growth stimulation of biological cells and tissue by electromagnetic fields and uses thereof |
JP2004290180A (en) * | 2003-03-10 | 2004-10-21 | Japan Science & Technology Agency | Magnetic field stimulation device, and method for promoting or inhibiting growth of living body cell or tissue using the magnetic field stimulating device |
-
2006
- 2006-06-22 BR BRPI0613092-5A patent/BRPI0613092A2/en not_active IP Right Cessation
- 2006-06-22 KR KR1020087002135A patent/KR20080023755A/en not_active Application Discontinuation
- 2006-06-22 MX MX2008000272A patent/MX2008000272A/en not_active Application Discontinuation
- 2006-06-22 WO PCT/US2006/024241 patent/WO2007005306A1/en active Application Filing
- 2006-06-22 JP JP2008519406A patent/JP2008544754A/en active Pending
- 2006-06-22 EP EP06785310A patent/EP1896566A1/en not_active Withdrawn
- 2006-06-22 CA CA002613408A patent/CA2613408A1/en not_active Abandoned
-
2007
- 2007-12-26 IL IL188443A patent/IL188443A0/en unknown
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2023110063A1 (en) * | 2021-12-14 | 2023-06-22 | Azyro Sa | Apparatus for in-vitro influencing biological cellular material, and use thereof |
Also Published As
Publication number | Publication date |
---|---|
KR20080023755A (en) | 2008-03-14 |
BRPI0613092A2 (en) | 2010-12-21 |
JP2008544754A (en) | 2008-12-11 |
IL188443A0 (en) | 2008-04-13 |
EP1896566A1 (en) | 2008-03-12 |
MX2008000272A (en) | 2008-03-11 |
WO2007005306A1 (en) | 2007-01-11 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP6380994B2 (en) | Alternating ion magnetic resonance (AIMR) multi-chamber culture apparatus and method of use thereof | |
US20070082328A1 (en) | Rotatable time varying electromagnetic force bioreactor and method of using the same | |
MX2014011369A (en) | Apparatus for local application of an extremely low frequency magnetic field to cells. | |
Smith et al. | Xenopus cell lines | |
Cheng et al. | Insulin-like growth factor-I and slow, bi-directional perfusion enhance the formation of tissue-engineered cardiac grafts | |
CA2613408A1 (en) | Time varying electromagnetic force sleeve for the expansion of cells and method of using the same | |
CN109402062A (en) | Application of the ZIP1 gene in the product that preparation inhibits apoptosis of mesenchymal stem cell | |
US20090220930A1 (en) | Time Varying Electromagnetic Force Sleeve for the Expansion of Cells and Method of Using the Same | |
JP6019404B2 (en) | Cell culture method, magnetic stimulation coil and power cable used in the method | |
US20080044890A1 (en) | Interchangable sleeve for enhancing proliferation of cells in a rotating bioreactor | |
CN108130275A (en) | A kind of Tissue Culture Dish for applying three-dimensional magnetic field | |
CN111411041A (en) | Gyromagnetic cell culture device and application thereof | |
US20060228795A1 (en) | Apparatus for enhancing proliferation of cells in a small-scale cell culturing container | |
CN207944108U (en) | A kind of Tissue Culture Dish applying three-dimensional magnetic field | |
JP2004290180A (en) | Magnetic field stimulation device, and method for promoting or inhibiting growth of living body cell or tissue using the magnetic field stimulating device | |
CN109385370A (en) | A kind of quick endothelialization device and method thereof of intravascular stent | |
Han et al. | Mouse embryonic tooth germ dissection and ex vivo culture protocol | |
CN112375679B (en) | Cell periodic pressure electric applying device and applying method | |
CN101213291A (en) | Time varying electromagnetic force sleeve for the expansion of cells and method of using the same | |
JP3554220B2 (en) | Fermentation method and fermentation apparatus | |
JP3836347B2 (en) | Method and apparatus for increasing the conductivity of water molecules | |
CN218755792U (en) | Three-dimensional cell culture device | |
US9096846B2 (en) | Separation of cultured cells | |
JP2008544754A5 (en) | ||
Schuetz et al. | Effects of time-varying magnetic fields on fibroblast growth |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
FZDE | Discontinued |