METHODS AND COMPOSITIONS FOR EXPANSION AND DIFFERENTIATION OF SKELETAL MUSCLE STEM CELLS OR PROGENITOR CELLS

• US 20180245048A1
• Filed: 02/25/2016
• Published: 08/30/2018
• Est. Priority Date: 02/25/2015
• Status: Abandoned Application

• Current Assignee: Agency For Science Technology and Research
• Original Assignee: Agency For Science Technology and Research
• Inventors: NG, Shyh Chang
• Application Number:
  US15/553,977
• Publication Number:
  US 20180245048A1
• Time in Patent Office: Days
• CPC Class Codes: C12N 2500/40 Nucleotides, nucleosides, bases cyclic nucleotides C12N2501/01, anti-neoplasic drugs C12N2501/06
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/415 Wnt; Frizzeled
• CPC Class Codes: C12N 2501/42 Notch; Delta; Jagged; Serrate
• CPC Class Codes: C12N 2501/999 Small molecules not provided for elsewhere
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 2506/45 from artificially induced pluripotent stem cells
• CPC Class Codes: C12N 5/0658 Skeletal muscle cells, e.g. myocytes, myotubes, myoblasts

Abstract

The present disclosure describes compositions for preparing a myofiber or myotube from a skeletal muscle stem cell or progenitor cell comprising a carnitine and/or a derivative thereof, a fatty acid a steroid and combinations thereof. Preferred embodiment comprises of 0.1 mM L-carnitine, 0.2 mM 9-cis-linoleic acid and 10 mM dihydrotestosterone. Also disclosed is a composition for inducing expansion of skeletal muscle stem cells or progenitor cells comprising a fibroblast growth factor agonist, a Notch signalling agonist, a nucleic acid, and combinations thereof. Preferred embodiment comprises 20 ng/ml basic fibroblast growth factor (bFGF), 50 μg/ml Delta-like ligand 1 (DLL1), 10 mM hypoxanthine and 1.6 mM thymidine.

Claims

1-38. -38. (canceled)

39. :

A composition for preparing a myofiber or myotube from a skeletal muscle stem cell or progenitor cell comprising a carnitine and/or a derivative thereof, a fatty add and a steroid.

40. :

The composition of claim 39, wherein the skeletal muscle progenitor cell is a myoblast cell; and

/or wherein the myoblast cell is a primary myoblast cell.

41. :

The composition of claim 39, wherein the skeletal muscle stem cell is a myosatellite cell.

42. :

The composition of claim 40, wherein the myoblast cell is derived from an embryonic stem (ES) cell, an induced pluripotent stem (IPS) cell, a mesenchymal stem cell, a neural stem cell or a multipotent stem cell; and

/or wherein the myoblast cell expresses high levels of Pax7 mRNA as compared to Pax7 mRNA expression in a human embryonic stem cell.

43. :

The composition of claim 39, wherein the skeletal muscle stem cell or progenitor cell is derived from a mammal; and

/or wherein the skeletal muscle stem cell or progenitor cell is derived from a human, rodent or primate.

44. :

The composition of claim 39, wherein the carnitine or derivative thereof is L-carnitine or an acyl carnitine; and

/or wherein the acyl carnitine is O-acetyl-carnitine, O-propionyl-carnitine or O-butanoyl-carnitine.

45. :

The composition of claim 39, wherein the fatty add is an unsaturated fatty add; and

/or wherein the fatty acid is an omega 3 or an omega 6 fatty acid.

46. :

The composition of claim 45, wherein the fatty add is selected from a group consisting of a linoleic acid, an arachidonic add, an eicosapentaenoic add, a docosahexaenoic acid, a linolenic acid and derivatives of the aforementioned fatty adds; and

/or wherein the linoleic acid is selected from a group consisting of 9-cis-linoleic acid, 12-cis-linoleic acid, cis-9 octadecadienoic acid and cis-12-octadecadienoic add.

47. :

The composition of claim 39, wherein the steroid is a sex steroid; and

/or wherein the sex steroid is selected from a group consisting of an androgen and estrogen; and

/or wherein the steroid is a testosterone, estradiol or a derivative thereof.

48. :

The composition of claim 39, wherein the composition comprises L-carnitine, linoleic acid and testosterone; and

/or wherein the composition comprises L-carnitine in a concentration of about 0.1 mM, linoleic add in a concentration of about 0.2 mM and testosterone in a concentration of about 10 mM.

49. :

A composition for inducing expansion of skeletal muscle stem cells or progenitor cells comprising a fibroblast growth factor signalling agonist, a Notch signalling agonist and a nucleic add.

50. :

The composition of claim 49, wherein the skeletal muscle progenitor cell is a myoblast cell, and/or wherein the myoblast cell is a primary myoblast cell.

51. :

The composition of claim 49, wherein the skeletal muscle stem cell is a myosatellite cell.

52. :

The composition of claim 50, wherein the myoblast cell is derived from an embryonic stem (ES) cell, an induced pluripotent stem (IPS) cell, a mesenchymal stem cell, a neural stem cell, or a multipotent stem cell; and

/or wherein the myoblast cell expresses high levels of Pax7 mRNA as compared to Pax7 mRNA expression in a human embryonic stem cell.

53. :

The composition of claim 49, wherein the skeletal muscle stem cell or progenitor cell is derived from a mammal; and

/or wherein the skeletal muscle stem cell or progenitor cell is derived from a human, rodent or primate.

54. :

The composition of claim 49, wherein the fibroblast growth factor signalling agonist is a fibroblast growth factor (FGF) and/or wherein the fibroblast growth factor signalling agonist is selected from a group consisting of FGF1, FGF2, FGF3, FGF4, FGF5, FGF6, FGF7, FGF8, FGF9, FGF10, FGF11, FGF12, FGF13, FGF14, FGF15, FGF16, FGF17, FGF18, FGF19, FGF20, FGF21, FGF22 and FGF23; and

/or wherein the fibroblast growth factor signalling receptor agonist is FGF2 (basic fibroblast growth factor;

bFGF) (bFGF) or a derivative thereof.

55. :

The composition of claim 49, wherein the Notch signalling agonist is a Delta-like ligand (DLL), a Jagged/Serrate ligand or a derivative thereof; and

/or wherein the Delta-like ligand (DLL) is selected from a group consisting of Delta-like 1 (DLL1), Delta-like 3 (DLL3) and Delta-like 4 (DLL4); and

/or wherein the Jagged/Serrate ligand is selected from a group consisting of Jagged 1 (JAG1), Jagged 2 (JAG2) and Serrate.

56. :

The composition of claim 49, wherein the nucleic add is derived from a nitrogenous base selected from a group consisting of hypoxanthine, adenine, guanine, thymine, cytosine, inosine, xanthine and a derivative of the aforementioned nitrogenous bases.

57. :

The composition of claim 49, wherein the composition comprises basic fibroblast growth factor (bFGF), Delta-like ligand 1 (DLL1), hypoxanthine and thymine; and

/or wherein the composition comprises basic fibroblast growth factor (bFGF) in a concentration of about 20 ng/ml, Delta-like ligand 1 (DLL1) in a concentration of about 50 μ

g/ml, hypoxanthine in a concentration of about 10 mM and thymine in a concentration of about 1.6 mM.

58. :

A method for preparing myofibers or myotubes, comprising contacting a skeletal muscle stem cell or progenitor cell with a composition for preparing a myofiber or myotube from a skeletal muscle stem cell or progenitor cell comprising a carnitine and/or a derivative thereof, a fatty add and a steroid.

59. :

A method for inducing expansion of skeletal muscle progenitor cells comprising contacting a skeletal muscle stem cell or progenitor cell with a composition for inducing expansion of skeletal muscle stem cells or progenitor cells comprising a fibroblast growth factor signalling agonist, a Notch signalling agonist and a nucleic add.