Isolation and preservation of fetal and neonatal hematopoietic stem and progenitor cells of the blood and methods of therapeutic use

• US 5,192,553 A
• Filed: 11/10/1988
• Issued: 03/09/1993
• Est. Priority Date: 11/12/1987
• Status: Expired due to Term

• Current Assignee: Pharmastem Therapeutics Incorporated
• Original Assignee: Biocyte Corporation
• Inventors: Broxmeyer, Hal E., Boyse, Edward A., Douglas, Gordon W.
• Primary Examiner(s): Rosen, Sam
• Application Number:
  US07/269,926
• Time in Patent Office: 1,580 Days
• Field of Search: 435/2, 435/172.1, 435/172.3, 435/240.2, 435/240.26
• US Class Current: 424/529
• 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 35/50 Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
• CPC Class Codes: A61K 35/51 Umbilical cord; Umbilical cord blood; Umbilical stem cells
• CPC Class Codes: C12N 2502/11 blood or immune system cells
• CPC Class Codes: C12N 2502/30 tumour cells
• CPC Class Codes: C12N 5/0647 Haematopoietic stem cells; Uncommitted or multipotent progenitors

Abstract

The present invention relates to hematopoietic stem and progenitor cells of neonatal or fetal blood that are cryopreserved, and the therapeutic uses of such stem and progenitor cells upon thawing. In particular, the present invention relates to the therapeutic use of fetal or neonatal stem cells for hematopoietic (or immune) reconstitution. Hematopoietic reconstitution with the cells of the invention can be valuable in the treatment or prevention of various diseases and disorders such as anemias, malignancies, autoimmune disorders, and various immune dysfunctions and deficiencies. In another embodiment, fetal or neonatal hematopoietic stem and progenitor cells which contain a heterologous gene sequence can be used for hematopoietic reconstitution in gene therapy. In a preferred embodiment of he invention, neonatal or fetal blood cells that have been cryopreserved and thawed can be used for utologous (self) reconstitution.

Claims

1. A method for obtaining human neonatal for fetal hematopoietic stem or progenitor cells comprising:

(a) isolating human neonatal or fetal blood components containing hematopoietic stem or progenitor cells;

(b) cryopreserving the blood components; and

(c) thawing the blood components,

such that the stem or progenitor cells are viable.

2. The method according to claim 1 further comprising the step after (c) of removing a cryopreservative.

3. The method according to claim 1 further comprising the step of growing the stem or progenitor cells in vitro.

4. The method according to claim 1 further comprising the step of enriching for stem and progenitor cells by a cell separation procedure.

5. The method according to claim 1 in which the blood components comprise whole blood.

6. The method according to claim 1 or 5 in which the blood components are isolated by collection from an umbilical cord.

7. The method according to claim 1 or 5 in which the blood components are isolated by collection from a placenta.

8. The method according to claim 1 or 5 in which the blood components are isolated by collection from both an umbilical cord and a placenta of the same individual.

9. The method according to claim 1 in which the cryopreservation is by use of a cryoprotective agent.

10. The method according to claim 9 in which the cryoprotective agent comprises dimethyl sulfoxide.

11. The method according to claim 1 in which the cryopreservation is by use of liquid nitrogen.

12. The method according to claim 9 in which the cryopreservation further comprises the use of liquid nitrogen.

13. A method for hematopoietic or immune reconstitution of a human comprising:

(a) isolating human neonatal or fetal blood components containing hematopoietic stem cells;

(b) cryopreserving the blood components;

(c) thawing the blood components; and

(d) introducing the blood components into a suitable human host,

such that the hematopoietic stem cells are viable and can proliferate within the host.

14. The method according to claim 13 in which the stem cells are autologous to the host.

15. The method according to claim 13 in which the stem cells are syngeneic to the host.

16. The method according to claim 13 in which the stem cells are allogeneic to the host.

17. The method according to claim 16 in which the host has Fanconi'"'"'s anemia.

18. The method according to claim 13 in which the blood components comprise whole blood.

19. The method according to claim 13 in which the blood components are isolated by collection from an umbilical cord.

20. The method according to claim 13 in which the blood components are isolated by collection from a placenta.

21. The method according to claim 13 in which the host is immunodeficient.

22. The method according to claim 21 in which the immunodeficiency is by reason of irradiation.

23. The method according to claim 21 in which the immunodeficiency is by reason of chemotherapy.

24. The method according to claim 21 in which the immunodeficiency is by reason of infection by a pathogenic microorganism.

25. The method according to claim 21 in which the host has a malignant solid tumor.

26. The method according to claim 13 in which the host has anemia.

27. The method according to claim 26 in which the host has Fanconi'"'"'s anemia.

28. The method according to claim 13 in which the host hyperproliferative stem cell disorder.

29. The method according to claim 13 in which the host has a hematopoietic malignancy.

30. The method according to claim 29 in which the hematopoietic malignancy is a leukemia.

31. The method according to claim 29 in which the hematopoietic malignancy is a lymphoma.

32. The method according to claim 13 in which the host has an autoimmune disease.

33. The method according to claim 13 in which the host has a hemolytic disorder.

34. The method according to claim 13 in which the host has a genetic disorder.

35. The method according to claim 34 in which the genetic disorder is Fanconi'"'"'s anemia.

36. The method according to claim 13 which further comprises, after step (a) or step (c), introducing a heterologous gene sequence into the stem cells, which gene sequence is stably incorporated and capable of expression by progeny of the stem cells.

37. The method according to claim 36 in which the host has a genetic disorder.

38. The method according to claim 37 in which the heterologous gene sequence comprises a sequence encoding hemoglobin.

39. The method according to claim 37 in which the host has thalassemia.

40. The method according to claim 37 in which the host has sickle cell disease.

41. The method according to claim 37 in which the host has anemia.

42. The method according to claim 36 in which the host is immunodeficient.

43. The method according to claim 42 in which the immunodeficiency is by reason of infection by a pathogenic microorganism.

44. The method according to claim 36 in which the host is infected by a pathogenic microorganism, and in which the heterologous gene sequence is expressed as a product which is toxic to the pathogenic microorganism without significant detriment to the host.

45. The method according to claim 36 in which the heterologous gene sequence is expressed as a nucleic acid sequence that is complementary to and can hybridize to a nucleic acid of a pathogenic microorganism.

46. The method according to claim 45 in which the pathogenic microorganism is Human Immunodeficiency Virus.

47. A method for hematopoietic or immune reconstitution of a human comprising:

(a) isolating human neonatal or fetal blood components containing hematopoietic stem and progenitor cells;

(b) cryopreserving the blood components;

(c) thawing the blood components; and

(d) introducing the blood components into a suitable human host,

such that the hematopoietic stem and progenitor cells are viable and can proliferate within the host.

48. The method according to claim 47 in which the stem and progenitor cells are autologous to the host.

49. The method according to claim 47 in which the stem and progenitor cells are syngeneic to the host.

50. The method according to claim 47 in which the stem and progenitor cells are allogeneic to the host.

51. The method according to claim 50 in which the host has Fanconi'"'"'s anemia.

52. The method according to claim 47 in which the blood components comprise whole blood.

53. The method according to claim 47 in which the blood components are isolated by collection from an umbilical cord.

54. The method according to claim 47 in which the blood components are isolated by collection from a placenta.

55. A method for obtaining cryopreserved human neonatal or fetal hematopoietic stem cells derived from the blood comprising:

(a) isolating human neonatal or fetal blood components containing human neonatal or fetal hematopoietic stem cells; and

(b) cryopreserving the blood components, such that the stem cells remain viable.

56. The method according to claim 55 in which the blood components comprise whole blood.

57. A method for hematopoietic or immune reconstitution of a human comprising introducing into the human a composition comprising human neonatal or fetal hematopoietic stem cells derived from the blood, in which the stem cells have been previously cryopreserved.

58. The method according to claim 57 in which the composition further comprises human neonatal or fetal hematopoietic progenitor cells derived from the blood, in which the progenitor cells have been previously cryopreserved.

59. The method according to claim 57 or 58 in which the composition comprises whole neonatal or fetal blood.

63. The method according to claim 57 in which a heterologous gene sequence is stably incorporated in the same cells.

64. The method according to claim 58 in which a heterologous gene sequence is stably incorporated in the progenitor cells.

60. A method for hematopoietic or immune reconstitution of a human comprising introducing into the human a composition comprising human neonatal or fetal hematopoietic stem cells derived from the blood, in which the stem cells are progeny of cells which have been previously cryopreserved.

61. The method according to claim 60 in which the composition further comprises human neonatal or fetal hematopoietic progenitor cells derived from the blood, in which the progenitor cells are progeny of cells which have been previously cryopreserved.

62. The method according to claim 60 or 61 in which the composition comprises whole neonatal or fetal blood.