EP0457856A4 - Isolation growth and differentiation of human muscle cells - Google Patents
Isolation growth and differentiation of human muscle cellsInfo
- Publication number
- EP0457856A4 EP0457856A4 EP19900910035 EP90910035A EP0457856A4 EP 0457856 A4 EP0457856 A4 EP 0457856A4 EP 19900910035 EP19900910035 EP 19900910035 EP 90910035 A EP90910035 A EP 90910035A EP 0457856 A4 EP0457856 A4 EP 0457856A4
- Authority
- EP
- European Patent Office
- Prior art keywords
- myoblasts
- muscle tissue
- dna construct
- myotubes
- cells
- 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.)
- Withdrawn
Links
Classifications
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- 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
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
- C12N15/86—Viral vectors
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- 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
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0658—Skeletal muscle cells, e.g. myocytes, myotubes, myoblasts
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
-
- 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
- C12N2500/00—Specific components of cell culture medium
- C12N2500/90—Serum-free medium, which may still contain naturally-sourced components
-
- 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
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/11—Epidermal growth factor [EGF]
-
- 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
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/30—Hormones
- C12N2501/33—Insulin
-
- 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
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/60—Transcription factors
-
- 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
- C12N2510/00—Genetically modified cells
-
- 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
- C12N2740/00—Reverse transcribing RNA viruses
- C12N2740/00011—Details
- C12N2740/10011—Retroviridae
- C12N2740/13011—Gammaretrovirus, e.g. murine leukeamia virus
- C12N2740/13041—Use of virus, viral particle or viral elements as a vector
- C12N2740/13043—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
Definitions
- the field of this invention is the development of myoblasts for use in the treatment of neuromuscular disease and for transformation for cellular therapy of diseases of diverse etiology.
- Myoblasts are precursor cells of the mesoderm that are destined for myogenesis.
- the determined myoblasts are capable of recognizing and spontaneously fusing with other myoblasts leading to the production of a differentiated myotube.
- the multinucleated myotube no longer divides or synthesizes DNA but produces muscle proteins in large quantity. These include constituents of the contractile apparatus and specialized cell-surface components essential to neuromuscular transmission.
- the differentiated muscle cell exhibits characteristic striations and rhythmic contractions.
- a further step in this pathway is maturation; the contractile apparatus and muscle at different stages of development contain distinct isoforms of muscle proteins such as myosin and actin, encoded by different members of multigene families.
- myoblasts have the potential for being used in a variety of ways.
- the myoblasts may serve for the treatment of various diseases associated with genetic defects involving muscle tissue.
- the myoblasts may also be found to be useful as vehicles for cell- therapy, where one or more genes may be introduced into the myoblasts to provide a product of interest.
- Blau and Webster, Proc. Natl. Acad. USA (1981) 75:5623-5627 describe isolation and cloning of muscle cells for proliferation or differentiation of individual clones.
- Blau, e_t a3.. , Proc. Natl. Acad. USA (1983) 80_:4856-4860 describe a defect in the proliferative capacity of myoblasts (satellite cells), mononucleated precursors of mature muscle fibers, in clonal analyses of cells cultured from Duchenne muscular dystrophy patients.
- Blau, et al. , Exp. Cell. Res. (1983) 144.495-503 describe the production and analysis of pure myoblast clones from biopsies of patients with Duchenne muscular dystrophy.
- Blau, e_t al. , Science (1985) 230;758-766 describe the fusion of muscle cells with non-muscle cells with activation of muscle gene expression in the non-muscle cell type.
- Webster, et al. , Exp. Cell Res. (1988) 174:252-265 describe the purification of human myoblasts using a fluorescence-activated cell sorter.
- Ham, e_t a ⁇ . , In Vitro Cell. & Dev. Biology (1988) 2 ⁇ 4:833-844 describe a serum-free medium for clonal growth of human muscle satellite cells.
- Myoblast cells are produced in serum free or low serum media for use in cell-therapy.
- the myoblasts are capable of migrating, fusing into pre-existing fibers, and may serve as carriers for genes introduced as a result of transformation.
- the migration of myoblasts across basal lamina allows for a reduced number of injections for treatment of a variety of diseases.
- Methods and cells are provided for use in cellular therapies for the treatment of diseases. Methods are described for preparing clonally pure or substantially enriched myoblasts in large amounts in the absence or substantial absence of serum in the nutrient medium. The resulting cells may be used for treatment of a variety of diseases associated with muscle tissue, or other tissue where a soluble factor is involved.
- the cells which are employed are myoblasts which may be obtained from tissue samples, which may include fetuses, neonates or tissue from older humans. These cells may be fresh or may be immediately frozen after being taken from the patient, or may be clonal cultures which are grown up from about 5 to 30 population doublings, and then stored frozen for use.
- the cells are grown in the subject media in a cell culture incubator (37°C, 5% C02 in air, saturated humidity), as the optimum conditions. Other conditions may be employed, if desired.
- the chosen medium provides for proliferation, without significant differentiation.
- the medium retains the myoblast level of maturation and, when desired, the myoblasts may be introduced into an environment, where they will differentiate and mature.
- the medium comprises a source of the essential amino acids, inorganic salts, trace elements and vitamins, as well as other organic components.
- the following table indicates what has generally been found to be optimum, although as is known in the field, various changes may be made to individual components without deleteriously affecting the growth of the myoblasts.
- Phenol Red (Na salt) 3.3 x 10 6 10 5 - 10 6 NaHC0 3 1.4 x 10 "2 5xl0 -2 - 5xl0 -3 VITAMINS d-Biotin 3.00 x 10 "8 10 -7 _ 10 -8
- Folinic Acid (Ca salt) «5H 2 0 1.00 x 10 -6 5x10 -6 _ 5x10 -7
- D-Pantothenic Acid (Hemi-Ca salt) 1.00 x 10 -4 5x10 -4 5x10 "
- Vitamin B12 1.00 x 10 ' ⁇ 8 5t.xv110n-8 _ 5x10 -9
- the first factor is dexamethasone in an amount of 0.3 to 0.5 ⁇ g/ml, preferably about 0.39 to 0.40 ⁇ g/ml.
- Serum albumin particularly bovine serum albumin, will be employed in from 0.25 to 0.75 mg/ml, preferably about 0.50 mg/ml.
- Epidermal growth factor is employed in from about 5 to 15 ng, preferably about 10 ng/ml.
- Fetuin is employed in from about 0.25 to 0.75 mg/ml, preferably 0.5 mg/ml.
- insulin conveniently bovine insulin, may be employed in from about 150 to 200 ⁇ g/ml, preferably about 180 ⁇ g/ml.
- an inoculum is introduced into the medium described above and the cells grown under the conditions described. After adding the inoculum, mild agitation is employed to ensure uniform distribution of the cells for further growth toward confluency.
- Tissue may be dissociated for a total of 40-60 min by two or three successive treatments with 0.05% trypsin - EDTA at 37°C in a Wheatin graduated trypsinzation flask with constant stirring.
- the cells collected in the supernatant after each trypsin treatment are pooled and cooled to 4°C on ice.
- Horse serum is added to a final concentration of 10% (vol/vol) to terminate further protease activity.
- the dissociated cells are then centrifuged (2 min, 25°C); the cell pellet is resuspended in conditioned media and either plated in culture or frozen in liquid nitrogen at a density of about 0.1 cm 3 of tissue per ml.
- the myoblasts may be transformed in any of a wide variety of ways, including fusion, transfection, infection, electroporation, ballistics or the like.
- the particular method for introducing the foreign DNA is not crucial to this invention.
- the gene of interest will be flanked by at least 50 bp on each side of DNA homologous to the site for integration, usually at least 100 bp, and the total of homologous DNA may be as high as 10 kbp, usually not greater than about 5 kbp, where preferably the flanking regions will be of about the same size.
- Regions for integration may include DNA sequences associated with a particular muscular defect.
- the host myoblasts may be removed from the host, transformed by homologous recombination, and cells cloned and screened for homologous recombination at the site of the defect.
- the myoblasts may then be used for expression of an endogenous gene (native to the host) or heterologous, which is normally not expressed in muscle tissue.
- cytokines for example, one may wish to provide for expression of cytokines, growth factors, colony stimulating factors, interferons, surface membrane receptors, insulin or the like.
- the myoblasts may provide for constitutive production of the expression product or alternatively or in combination, one may introduce a receptor for the soluble product, which provides for inducible transcription of a cellular, e.g., cytoplasmic, nuclear, etc., protein. By activating the receptor, the myoblasts may be induced to produce the expression product under the- induction of the relevant ligand.
- a receptor for the soluble product which provides for inducible transcription of a cellular, e.g., cytoplasmic, nuclear, etc., protein.
- the myoblasts may be induced to produce the expression product under the- induction of the relevant ligand.
- Various vehicles or vector constructs may be employed for the transformation of the myoblast cells. Of particular interest for transfection or infection are replication-defective viral vectors, DNA virus or retroviral vectors, which may be introduced into the cells.
- the vectors will normally be free of any prokaryotic DNA and may comprise a number of different functional sequences.
- one of the functional sequences may be a DNA region comprising transcriptional and translational initiation and termination regulatory sequences, an open reading frame encoding the protein of interest, and may further comprise flanking regions for site directed integration.
- the 5 '-flanking region will provide for homologous recombination to change the nature of the transcriptional initiation region.
- the presence or absence of an enhancer may be modified, to provide for inducible transcription or non-inducible transcription, to increase or decrease the level of transcription, or the like.
- the promoter region may be modified, so as to be more or less susceptible to induction, to increase or decrease the level of transcription, or the like.
- the structural gene which is employed may result in an intracellular product, i.e., retained in the cell, in the cytoplasm or organelle, e.g., the nucleus, in transport to a membrane, either an intracellular membrane or the cell membrane, or for secretion by providing for the natural signal sequence present with the structural gene or a signal sequence which is not naturally present with the structural gene.
- the soluble protein of interest is a fragment of a larger protein, it may be necessary to provide a signal sequence with such protein, so that upon secretion and processing at the processing site, the desired protein will have the natural sequence.
- a marker may be present for selection of cells which contain the vehicle construct. Normally, the marker will allow for positive selection, in providing protection from one or more cytotoxic agents. For example, kanamycin resistance may be employed, where the cells may be selected with G418, dihydrofolate reductase may be employed for resistance to methotrexate, and the like.
- the marker may be an inducible or non-inducible gene, so that selection may occur under induction or without induction.
- the vector may also include a replication origin and such other genes which are necessary for replication in the host.
- the replication system comprising the origin and any proteins associated with replication encoded by the particular virus may be included as part of a construct. Care must be taken in selecting the replication system, so that the genes which are encoded for replication do not provide for transformation of the myoblasts.
- Illustrative replication systems include Epstein-Barr virus.
- replication defective vehicles may be employed, particularly replication-defective retroviral vectors. These vectors are described by Price, e_t al. , Proc. Natl. Acad. Sci.(1987) 84:156-160 and Sares, et al. EMBO J. (1986) 5:3133-3142.
- the final vehicle construct may have one or more genes of interest. Either a cDNA gene or a chromosomal gene may be employed. Of particular interest is to provide for at least one intron, which may be present in the 5 '-non- coding region or in the coding region. It is found that the presence of an intron enhances stability of the messenger RNA.
- cells may be transformed in vivo by injection of replication-defective viral vectors, which are infectious.
- the vectors may be introduced into retroviral producer cells for ecotropic packaging.
- the cells are then collected, filtered and concentrated by centrifugation and the viral stock may then be injected into a site ij vivo. Since it is found that the myoblasts will migrate, relatively few injections into the muscle fibers are required, since the myoblasts will expand into adjacent regions.
- the vector is conveniently by injection in a physiologically acceptable medium, such as water, saline, phosphate buffered saline, or the like.
- a physiologically acceptable medium such as water, saline, phosphate buffered saline, or the like.
- the viral concentration will generally be from about 10 5 ffu.
- Other additives which may be present include polybrene.
- the injections will be about 10 5 cells per cm 3 of muscle tissue.
- the trauma to the tissue may be substantially minimized by having only a few injections in the region of interest. Particularly, where a patient may have need for extensive treatment, the desirability of having a low number of injections in a particular area is manifest.
- Two ⁇ -galactosidase vectors were employed, referred to as BAG and pMMuLVSVnlsLacZ.
- the two vectors each contain ⁇ -galactosidase encoding sequences under the transcriptional control of the MMuLV promoter/enhancer or the SV40 early promoter.
- the vectors are further characterized by neo r and LacZ genes, and in some cases 7 amino acid codons for the SV40 large T nuclear localization sequences.
- Clusters of muscle fibers stained blue were scattered throughout the lower hind limb. Charcoal particles were generally located between soleus and lateral gastrocnemius and were used to identify the site of infection. No differences in the distribution or size of clusters of labeled muscle cells was observed with either the BAG or pMMuLVSVnlsLacZ vectors.
- clusters of labeled cells close to the charcoal particles were not detectably different from those several millimeters away, suggesting that the injection did not perturb development of nearby tissue.
- Clusters of stained muscle fibers in the lateral gastrocnemius muscle demonstrated that the myoblasts could be infected and expressed ⁇ -galacto- sidase even after fusion into multinucleate fibers.
- Many of the clusters observed represented clones derived from single cells, some of the progeny of which migrated across basal lamina of a given muscle fiber and fused into adjacent muscle fibers. Since each myoblast is associated with a single fiber at the time of infection, the data indicate that myoblasts are capable of migrating through the basal lamina from one fiber to another.
- migration appears to be a relatively frequent event.
- the subject invention allows for the use of muscle forming cells in the treatment of diseases associated with muscle tissue or for production of soluble or other proteins in a host.
- the myoblasts are capable of forming new muscle tissue or participating in the formation of fibers, where the cells may provide for useful properties, correct defects, and the like.
- the cells may be modified with markers, to allow for selective advantage of the transformed cells over the naturally present cells.
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- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Rheumatology (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Public Health (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Cell Biology (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Veterinary Medicine (AREA)
- Virology (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
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Abstract
Description
Claims
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US36537489A | 1989-06-13 | 1989-06-13 | |
US365374 | 2003-02-11 |
Publications (2)
Publication Number | Publication Date |
---|---|
EP0457856A1 EP0457856A1 (en) | 1991-11-27 |
EP0457856A4 true EP0457856A4 (en) | 1992-10-21 |
Family
ID=23438629
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP19900910035 Withdrawn EP0457856A4 (en) | 1989-06-13 | 1990-06-13 | Isolation growth and differentiation of human muscle cells |
Country Status (4)
Country | Link |
---|---|
EP (1) | EP0457856A4 (en) |
JP (1) | JP3244696B2 (en) |
CA (1) | CA2058955A1 (en) |
WO (1) | WO1990015863A1 (en) |
Families Citing this family (23)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6048729A (en) | 1987-05-01 | 2000-04-11 | Transkaryotic Therapies, Inc. | In vivo protein production and delivery system for gene therapy |
WO1990006997A1 (en) * | 1988-12-13 | 1990-06-28 | United States Government As Represented By The Secretary Of The Department Of Health And Human Services | Genetically engineered endothelial cells and use thereof |
US5538722A (en) * | 1989-06-13 | 1996-07-23 | Stanford University | Isolation, growth, differentiation and genetic engineering of human muscle cells |
WO1991012329A2 (en) * | 1990-02-12 | 1991-08-22 | Board Of Regents, The University Of Texas System | Satellite cell proliferation in adult skeletal muscle |
US5352595A (en) * | 1991-09-03 | 1994-10-04 | Fred Hutchinson Cancer Research Center | Myod regulatory region |
US6063630A (en) | 1991-11-05 | 2000-05-16 | Transkaryotic Therapies, Inc. | Targeted introduction of DNA into primary or secondary cells and their use for gene therapy |
PT101031B (en) * | 1991-11-05 | 2002-07-31 | Transkaryotic Therapies Inc | PROCESS FOR THE SUPPLY OF PROTEINS BY GENETIC THERAPY |
US6692737B1 (en) | 1991-11-05 | 2004-02-17 | Transkaryotic Therapies, Inc. | In vivo protein production and delivery system for gene therapy |
US6054288A (en) * | 1991-11-05 | 2000-04-25 | Transkaryotic Therapies, Inc. | In vivo protein production and delivery system for gene therapy |
US6348327B1 (en) | 1991-12-06 | 2002-02-19 | Genentech, Inc. | Non-endocrine animal host cells capable of expressing variant proinsulin and processing the same to form active, mature insulin and methods of culturing such cells |
JPH07509217A (en) * | 1992-03-04 | 1995-10-12 | ウォルフ ジョン エイ | How to deliver therapeutic substances to the brain |
JPH08501210A (en) * | 1992-07-02 | 1996-02-13 | ザ ソールク インスチチュート フォア バイオロジカル スタディズ | Utilization of myoblasts for sustained release of gene products |
US6531124B1 (en) | 1992-07-10 | 2003-03-11 | Transkaryotic Therapies, Inc. | In vivo production and delivery of insulinotropin for gene therapy |
US6670178B1 (en) | 1992-07-10 | 2003-12-30 | Transkaryotic Therapies, Inc. | In Vivo production and delivery of insulinotropin for gene therapy |
US5935849A (en) | 1994-07-20 | 1999-08-10 | Cytotherapeutics, Inc. | Methods and compositions of growth control for cells encapsulated within bioartificial organs |
US5858747A (en) * | 1994-07-20 | 1999-01-12 | Cytotherapeutics, Inc. | Control of cell growth in a bioartificial organ with extracellular matrix coated microcarriers |
GB9419048D0 (en) * | 1994-09-20 | 1994-11-09 | Watt Diana J | Treatment of muscular disorders |
US6495364B2 (en) * | 1995-05-23 | 2002-12-17 | Neurotech, S.A. | Mx-1 conditionally immortalized cells |
EP1200559A2 (en) | 1999-07-23 | 2002-05-02 | Diacrin, Inc. | Muscle cells and their use in cardiac repair |
US8889122B2 (en) | 2005-05-09 | 2014-11-18 | Mytogen, Inc. | Cellular cardiomyoplasty as supportive therapy in patients with heart disease |
WO2010031190A1 (en) * | 2008-09-22 | 2010-03-25 | UNIVERSITé LAVAL | Culture medium for myoblasts, precursors thereof and derivatives thereof |
FR2960783B1 (en) | 2010-06-04 | 2012-07-27 | Ass Pour Les Transferts De Technologies Du Mans | FUNCTIONALIZED MEMBRANE FOR ENCAPSULATING CHAMBER OF CELLS PRODUCING AT LEAST ONE SUBSTANCE OF THERAPEUTIC INTEREST AND BIOARTIFICIAL ORGAN COMPRISING SUCH A MEMBRANE |
EP3095509A1 (en) | 2015-05-18 | 2016-11-23 | Defymed | Membranes functionalized with heparin for bioartificial organs |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1988008450A1 (en) * | 1987-05-01 | 1988-11-03 | Birdwell Finlayson | Gene therapy for metabolite disorders |
WO1989002468A1 (en) * | 1987-09-11 | 1989-03-23 | Whitehead Institute For Biomedical Research | Transduced fibroblasts and uses therefor |
-
1990
- 1990-06-13 WO PCT/US1990/003352 patent/WO1990015863A1/en not_active Application Discontinuation
- 1990-06-13 JP JP50911490A patent/JP3244696B2/en not_active Expired - Fee Related
- 1990-06-13 EP EP19900910035 patent/EP0457856A4/en not_active Withdrawn
- 1990-06-13 CA CA002058955A patent/CA2058955A1/en not_active Abandoned
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1988008450A1 (en) * | 1987-05-01 | 1988-11-03 | Birdwell Finlayson | Gene therapy for metabolite disorders |
WO1989002468A1 (en) * | 1987-09-11 | 1989-03-23 | Whitehead Institute For Biomedical Research | Transduced fibroblasts and uses therefor |
Non-Patent Citations (3)
Title |
---|
JOURNAL OF CELLULAR BIOCHEMISTRY vol. SUPPL, no. 14E, 31 March 1990, page 231; BLAU, H. M. ET AL.: 'Influence of immune response and migration on myoblast cell therapy' * |
MOLECULAR AND CELLULAR BIOLOGY vol. 10, no. 6, 1 June 1990, pages 3268 - 3271; SMITH, B. F. ET AL.: 'Genes transferred by retroviral vectors into normal and mutant myoblasts in primary cultures are expressed in myotubes' * |
See also references of WO9015863A1 * |
Also Published As
Publication number | Publication date |
---|---|
WO1990015863A1 (en) | 1990-12-27 |
JPH05500601A (en) | 1993-02-12 |
EP0457856A1 (en) | 1991-11-27 |
CA2058955A1 (en) | 1990-12-14 |
JP3244696B2 (en) | 2002-01-07 |
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