Methods for obtaining stem cells

• US 10,076,113 B2
• Filed: 10/11/2016
• Issued: 09/18/2018
• 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): Berke-Schlessel, David W
• Application Number:
  US15/290,825
• Publication Number:
  US 20170086453A1
• Time in Patent Office: 707 Days
• Field of Search: None
• 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 mammalian stem cells comprising:

a) maintaining a biological sample comprising mammalian stem cells in a culture 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; and

b) selecting viable stem cells.

2. The method according to claim 1, wherein the biological material is obtained from a human.

3. The method according to claim 1, wherein the biological material is bone marrow.

4. The method according to claim 1, wherein the biological material is human bone marrow.

5. The method according to claim 1, wherein the biological material is obtained from a tissue or organ selected from muscle, central nervous system, peripheral blood, umbilical cordon cord blood, skin epidermis, dental pulp, heart, gut, pancreas, lung, adipose, ovarian epithelium, retina, cornea, or testis.

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

C. within 48 hours after death.

7. The method according to claim 1, further comprising resuspending the selected viable stem cells in a pharmaceutically acceptable carrier.

8. The method according to claim 1, wherein the biological material comprising mammalian stem cells is maintained at an oxygen concentration equal to or less than 0.1% and at a temperature of 1-6°

C. for at least 4 days.

9. A method of obtaining mammalian stem cells expressing a transgene of interest comprising:

a) maintaining a biological material comprising mammalian stem cells in a culture 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;

b) selecting viable stem cells; and

c) transforming or transfecting the viable stem cells with a transgene of interest.

10. The method according to claim 9, wherein the biological material is obtained from a human.

11. The method according to claim 9, wherein the biological material is bone marrow.

12. The method according to claim 9, wherein the biological material is human bone marrow.

13. The method according to claim 9, wherein the biological material is obtained from a tissue or organ selected from muscle, central nervous system, peripheral blood, umbilical cord blood, skin epidermis, dental pulp, heart, gut, pancreas, lung, adipose, ovarian epithelium, retina, cornea, or testis.

14. The method according claim 9, wherein the biological material is obtained from a cadaver stored at 1-6°

C. within 48 hours after death.

15. The method according to claim 9, further comprising resuspending the transformed or transfected stem cells in a pharmaceutically acceptable carrier.

16. The method according to claim 9, wherein the biological material comprising mammalian stem cells is maintained at an oxygen concentration equal to or less than 0.1% and at a temperature of 1-6°

C. for at least 4 days.

17. A method of obtaining mammalian stem cells comprising:

a) providing a culture comprising mammalian stem cells;

b) removing oxygen from the culture to create an anoxic culture;

c) maintaining the culture in anoxic conditions at a temperature of 1-6°

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

d) selecting viable stem cells.

18. The method according to claim 17, wherein the biological material is obtained from a human.

19. The method according to claim 17, wherein the biological material is bone marrow.

20. The method according to claim 17, wherein the biological material is human bone marrow.

21. The method according to claim 17, wherein the biological material is obtained from a tissue or organ selected from muscle, central nervous system, peripheral blood, umbilical cord blood, skin epidermis, dental pulp, heart, gut, pancreas, lung, adipose, ovarian epithelium, retina, cornea, or testis.

22. The method according claim 17, wherein the biological material is obtained from a cadaver stored at 1-6°

C. within 48 hours after death.

23. The method according to claim 17, further comprising resuspending the selected viable stem cells in a pharmaceutically acceptable carrier.

24. The method according to claim 17, wherein the biological material comprising mammalian stem cells is maintained at an oxygen concentration equal to or less than 0.1% and at a temperature of 1-6°

C. for at least 4 days.