CA2269096A1 - Method for in vitro preconditioning of myoblasts before transplantation - Google Patents

Method for in vitro preconditioning of myoblasts before transplantation

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Publication number
CA2269096A1
CA2269096A1 CA002269096A CA2269096A CA2269096A1 CA 2269096 A1 CA2269096 A1 CA 2269096A1 CA 002269096 A CA002269096 A CA 002269096A CA 2269096 A CA2269096 A CA 2269096A CA 2269096 A1 CA2269096 A1 CA 2269096A1
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CA
Canada
Prior art keywords
donor
myoblasts
functional
cells
recombinant
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.)
Granted
Application number
CA002269096A
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French (fr)
Other versions
CA2269096C (en
Inventor
Jacques P. Tremblay
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Universite Laval
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Individual
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Application filed by Individual filed Critical Individual
Publication of CA2269096A1 publication Critical patent/CA2269096A1/en
Application granted granted Critical
Publication of CA2269096C publication Critical patent/CA2269096C/en
Anticipated expiration legal-status Critical
Expired - Fee Related legal-status Critical Current

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Classifications

    • 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
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • 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
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/01—Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin
    • 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
    • C12N2501/115—Basic fibroblast growth factor (bFGF, FGF-2)
    • 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
    • C12N2501/50—Cell markers; Cell surface determinants
    • C12N2501/59—Lectins
    • 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
    • C12N2501/70—Enzymes
    • 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
    • C12N2502/00—Coculture with; Conditioned medium produced by
    • C12N2502/13—Coculture with; Conditioned medium produced by connective tissue cells; generic mesenchyme cells, e.g. so-called "embryonic fibroblasts"
    • C12N2502/1323—Adult fibroblasts
    • 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

Abstract

A method of pretreating healthy donor's myoblast cultures with growth or trophic factors like basic fibroblast growth factor (bFGF) and with concanavalin A on transplantation to subjects suffering of myopathy like muscular dystrophy is disclosed and claimed. Recipient muscles show a higher percentage of functional cells, a four-fold increase, demonstrated by the higher incidence of dystrophin-positive fibers, and does not require previous preconditioning of recipient muscles by irradiation or toxin administration. The recipient subjects were immunosuppressed with FK 506. When growing myoblasts with 20 µg/ml concanavalin A or 100 ng/ml TPA for two to four days, migration of donor cells in recipient tissue was increased by 3 - 4 fold. This suggests that, when using primary cultures, metalloproteases are secreted by fibroblasts, resulting in a greater degradation of the extracellular matrix. Both metalloproteases and bFGF appear beneficial for the success of the transplantation. The use of recombinant myoblast expressing metalloproteases is also contemplated.

Claims (17)

1. A method for increasing the number of transplanted functional donor's myoblasts which are fused with non-functional myoblasts of a recipient individual suffering of a myopathy, which comprises the step of growing in vitro said donor's myoblasts in a appropriate culture medium in the presence of fibroblasts and of an agent inducing the secretion of an enzyme involved in extracellular matrix destruction, prior to injecting a mixture comprising said donor's myoblasts and induced enzyme into said recipient individual's muscle, whereby a functional muscle is at least in part restored.
2. A method for increasing the number of transplanted functional donor's myoblasts which are fused with non-functional myoblasts of a recipient individual suffering of a myopathy, which comprises the steps of: inserting into said donor's myoblasts a gene construct capable of expressing an enzyme involved in extracellular matrix destruction, obtaining thereby recombinant donor's myoblasts, and growing said recombinant donor's myoblasts in an appropriate culture medium, prior to injecting said recombinant donor's myoblasts into said recipient individual's muscle, whereby a functional muscle is at least in part restored.
3. A method of claim 1, wherein said enzyme is a metalloprotease.
4. A method of claim 2, wherein said enzyme is a metalloprotease.
5. A method of claim 3, wherein said metalloprotease is Gelatinase A or Matrilysine.
6. A method of claim 4, wherein said metalloprotease is Gelatinase A or Matrilysine.
7. A method of claim 1, 3 or 5, wherein said agent is Concanavalin A or phorbol ester.
8. A method as defined in any one of claims 1 to 7, wherein said culture medium further comprises a growth or trophic factor for increasing the multiplication of said donor's myoblasts.
9. A method as defined in claim 8, wherein said growth or trophic factor is selected from the group consisting of basic fibroblast growth factor (bFGF), insulin growth factor I, transferrin, platelet-derived growth factor, epidermal growth factor, adrenocorticotrophin, macrophage colony-stimulating factor, protein kinase C activators, agonists thereof, and combinations thereof.
10. A method as defined in claim 9, wherein said factor is bFGF.
11. A method as defined in any one of claims 1, 3, 5, 7, 8, 9, and 10, wherein said donor's myoblasts are obtained for a primary myoblast culture resulting from culturing a cell dispersion of donor's muscle biopsy.
12. A method as defined in claim 11, wherein said primary myoblast culture is grown in the presence of 100 ng of recombinant human basic fibroblast growth factor per milliliter of culture medium for a period of time of about 48 hours before transplantation.
13. A method as defined in claim 11, wherein said primary myoblast culture is grown in the presence of 100 ng of recombinant human basic fibroblast growth factor and 20 µg Concanavalin A per milliter of culture medium for a period of time of about 48 hours before transplantation.
14. A method as defined in any one of claims 1 to 13, wherein said myopathy is Duchenne muscular dystrophy.
15. A method for increasing the number of transplanted functional donor's cells which are fused with corresponding non-functional cells of a recipient individual's tissue, which comprises the steps of:
inserting into said donor's cells a gene construct capable of expressing an enzyme involved in extracellular matrix destruction, obtaining thereby recombinant donor's cells, and growing said recombinant donor's cells in an appropriate culture medium, prior to injecting said recombinant donor's cells into said recipient individual's tissue, whereby a functional tissue is at least in part restored.
16. A method of claim 15, wherein said enzyme is a metalloprotease.
17. A method of claim 16, wherein said metalloprotease is Matrilysine or Gelatinase A.
CA002269096A 1996-10-18 1997-10-17 Method for in vitro preconditioning of myoblasts before transplantation Expired - Fee Related CA2269096C (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US2869296P 1996-10-18 1996-10-18
US60/028,692 1996-10-18
PCT/CA1997/000774 WO1998017784A1 (en) 1996-10-18 1997-10-17 Method for in vitro preconditioning of myoblasts before transplantation

Publications (2)

Publication Number Publication Date
CA2269096A1 true CA2269096A1 (en) 1998-04-30
CA2269096C CA2269096C (en) 2009-12-15

Family

ID=21844898

Family Applications (1)

Application Number Title Priority Date Filing Date
CA002269096A Expired - Fee Related CA2269096C (en) 1996-10-18 1997-10-17 Method for in vitro preconditioning of myoblasts before transplantation

Country Status (7)

Country Link
US (3) US20020012657A1 (en)
EP (1) EP0946713B1 (en)
AT (1) ATE356199T1 (en)
AU (1) AU4613397A (en)
CA (1) CA2269096C (en)
DE (1) DE69737460T2 (en)
WO (1) WO1998017784A1 (en)

Families Citing this family (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
IT1299201B1 (en) * 1998-05-08 2000-02-29 San Raffaele Centro Fond GENETICALLY MODIFIED FIBROBLASTS AND THEIR USE
US6284242B1 (en) * 1999-04-16 2001-09-04 Regents Of The University Of Michigan Method for enhancing myoblast migration and invasion in the context of gene therapy
US6673604B1 (en) 1999-07-23 2004-01-06 Diacrin, Inc. Muscle cells and their use in cardiac repair
US20030113301A1 (en) * 1999-07-23 2003-06-19 Albert Edge Muscle cells and their use in cardiac repair
US7627373B2 (en) * 2002-11-30 2009-12-01 Cardiac Pacemakers, Inc. Method and apparatus for cell and electrical therapy of living tissue
US20040158289A1 (en) * 2002-11-30 2004-08-12 Girouard Steven D. Method and apparatus for cell and electrical therapy of living tissue
US7840263B2 (en) * 2004-02-27 2010-11-23 Cardiac Pacemakers, Inc. Method and apparatus for device controlled gene expression
US7764995B2 (en) 2004-06-07 2010-07-27 Cardiac Pacemakers, Inc. Method and apparatus to modulate cellular regeneration post myocardial infarct
US7828711B2 (en) * 2004-08-16 2010-11-09 Cardiac Pacemakers, Inc. Method and apparatus for modulating cellular growth and regeneration using ventricular assist device
US8060219B2 (en) * 2004-12-20 2011-11-15 Cardiac Pacemakers, Inc. Epicardial patch including isolated extracellular matrix with pacing electrodes
US7981065B2 (en) 2004-12-20 2011-07-19 Cardiac Pacemakers, Inc. Lead electrode incorporating extracellular matrix
JP2008537942A (en) * 2005-03-31 2008-10-02 マイトジェン， インコーポレイテッド Treatment for heart disease
US8889122B2 (en) 2005-05-09 2014-11-18 Mytogen, Inc. Cellular cardiomyoplasty as supportive therapy in patients with heart disease
AU2017351638A1 (en) 2016-10-26 2019-06-13 Genea Biocells USA (Holdings), Inc. Improved generation of muscle lineage cells and therapeutic uses thereof

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU624284B2 (en) * 1989-11-24 1992-06-04 Monash University Proliferative action of leukaemia inhibitory factor on satellite cells
AU7312891A (en) * 1990-02-12 1991-09-03 Board Of Regents, The University Of Texas System Satellite cell proliferation in adult skeletal muscle
US5833978A (en) * 1995-03-16 1998-11-10 Universite Laval Method of in vitro preconditioning healthy donor's myoblasts before transplantation thereof in compatible patients suffering of recessive myopathies like muscular dystrophy, for improving transplantation success
US6284242B1 (en) 1999-04-16 2001-09-04 Regents Of The University Of Michigan Method for enhancing myoblast migration and invasion in the context of gene therapy

Also Published As

Publication number Publication date
EP0946713A1 (en) 1999-10-06
ATE356199T1 (en) 2007-03-15
US20070178077A1 (en) 2007-08-02
EP0946713B1 (en) 2007-03-07
WO1998017784A1 (en) 1998-04-30
US20020012657A1 (en) 2002-01-31
US7189391B2 (en) 2007-03-13
CA2269096C (en) 2009-12-15
AU4613397A (en) 1998-05-15
DE69737460T2 (en) 2008-04-10
US20020182193A1 (en) 2002-12-05
DE69737460D1 (en) 2007-04-19

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