Methods for obtaining stem cells

• US 9,499,792 B2
• Filed: 06/03/2011
• Issued: 11/22/2016
• Est. Priority Date: 06/04/2010
• Status: Active Grant

• Current Assignee: Centre National De La Recherche Scientifique, Institut Pasteur
• Original Assignee: Centre National De La Recherche Scientifique, Institut Pasteur
• Inventors: Chretien, Fabrice, Latil, Mathilde, Tajbakhsh, Shahragim
• Primary Examiner(s): Claytor, Renee
• Assistant Examiner(s): BERKE-SCHLESSEL, DAVID W
• Application Number:
  US13/700,832
• Publication Number:
  US 20130149286A1
• Time in Patent Office: 1,999 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A01N 1/021 Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
• CPC Class Codes: A01N 1/0284 Temperature processes, i.e. using a designated change in temperature over time
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: A61K 35/14 Blood; Artificial blood perfluorocarbons A61K31/02; umbilical cord blood A61K35/51; haemoglobin A61K38/42
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61K 35/34 Muscles; Smooth muscle cells; Heart; Cardiac stem cells; Myoblasts; Myocytes; Cardiomyocytes vascular smooth muscle A61K35/44
• CPC Class Codes: C12N 2500/02 Atmosphere, e.g. low oxygen conditions
• CPC Class Codes: C12N 5/0623 Stem cells
• CPC Class Codes: C12N 5/0647 Haematopoietic stem cells; Uncommitted or multipotent progenitors
• CPC Class Codes: C12N 5/0659 Satellite cells
• CPC Class Codes: C12N 5/0668 Mesenchymal stem cells from other natural sources

Abstract

The present invention relates to methods for obtaining stem cells from mammalian cadavers, methods for obtaining or purifying stem cells from a sample likely to contain non-stem cells, methods of regeneration of injured tissues and methods of treatment.

Claims

1. A method of obtaining skeletal muscle stem cells comprising the following steps:

a) maintaining a biological sample at an oxygen concentration equal to or less than 0.1% and at a temperature of 1-6°

C. during a period of time of at least 2 days wherein the biological material is a skeletal muscle sample; and

b) selecting the viable cells, wherein viable cells are skeletal muscle cells.

3. The method according to claim 1, wherein the biological material is obtained from a living donor, except human embryo or foetus.

4. The method according to claim 1, wherein the biological material is maintained at 1-6°

C. during a period of time of 2 days to 60 days.

5. The method according to claim 1, wherein the biological material is maintained at 1-6°

C. in the absence of oxygen.

6. The method according claim 1, wherein the biological material was obtained from a mammal cadaver stored at 1-6°

C. within 48 hours after death.

7. The method according to claim 3, wherein the biological material is obtained from a living mammal.

8. The method according to claim 1, further comprising an ultimate step following step b) consisting of resuspending the selected viable skeletal muscle stem cells in a pharmaceutically acceptable carrier.

11. The method according to claim 1, wherein the skeletal muscle sample is maintained at an oxygen concentration equal to or less than 0.1% and at a temperature of 1-6°

C. during a period of time of at least 4 days.

2. A method of obtaining mammalian skeletal muscle stem cells expressing a transgene of interest, comprising the following steps:

a) maintaining a biological material at an oxygen concentration equal to or less than 0.1% and at a temperature of 1-6°

C. during a period of time of at least 2 days, wherein the biological material is a skeletal muscle sample;

b) selecting viable cells, wherein viable cells are skeletal muscle stem cells;

c) transforming or transfecting the viable skeletal muscle stem cells.

10. The method according to claim 2, further comprising an ultimate step following step c) consisting of resuspending the selected viable skeletal muscle stem cells in a pharmaceutically acceptable carrier.

12. The method according to claim 2, wherein the skeletal muscle sample is maintained at an oxygen concentration equal to or less than 0.1% and at a temperature of 1-6°

C. during a period of time of at least 4 days.

9. A method for selecting skeletal muscle stem cells comprising using an anaerobic cell culture system for selecting stem cells by maintaining a skeletal muscle sample comprising stem cells and non-stem cells at an oxygen concentration equal to or less than 0.1% and at a temperature of 1-6°

C. during a period of time of at least 2 days.

13. The method according to claim 9, wherein the skeletal muscle sample is maintained at an oxygen concentration equal to or less than 0.1% and at a temperature of 1-6°

C. during a period of time of at least 4 day.