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

• US 6,461,645 B1
• Filed: 05/16/1990
• Issued: 10/08/2002
• Est. Priority Date: 11/12/1987
• Status: Expired due to Fees

• Current Assignee: Pharmastem Therapeutics Incorporated
• Original Assignee: Pharmastem Therapeutics Incorporated
• Inventors: Douglas, Gordon W., Broxmeyer, Hal E., Boyse, Edward A.
• Primary Examiner(s): Witz, Jean C.
• Application Number:
  US07/525,428
• Time in Patent Office: 4,528 Days
• Field of Search: 435/2, 424/529, 424/937
• 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 the invention, neonatal or fetal blood cells that have been cryopreserved and thawed can be used for autologous (self) reconstitution.

Claims

1. A cryopreserved pharmaceutical composition comprising:

(a) viable human neonatal or fetal hematopoietic stem cells derived from the umbilical cord blood or placental blood of a single human collected at the birth of said human, in which said cells are present in an amount sufficient to effect hematopoietic reconstitution of a human adult;

(b) an amount of cryopreservative sufficient for cryopreservation of said cells; and

(c) a pharmaceutically acceptable carrier.

2. The composition of claim 1 which further comprises a viable human neonatal or fetal hematopoietic progenitor cell.

3. The composition of claim 1 which comprises whole neonatal or fetal blood.

4. The composition of claim 1 which further comprises an anticoagulent.

5. The composition of claim 1, 2, 3 or 4 in which the cryopreservative comprises dimethyl sulfoxide.

6. A pharmaceutical composition comprising a pharmaceutically acceptable carrier;

and human neonatal or fetal hematopoietic stem cells obtained by a method comprising the following steps in the stated order;

(a) isolating human neonatal or fetal blood components containing hematopoietic stem cells derived from the umbilical cord blood or placental blood of a single human at the birth of said human, in which said cells are present in an amount sufficient to effect hematopoietic reconstitution of a human adult;

(b) cryopreserving the blood components in the presence of a cryopreservative with an amount of a cryopreservative sufficient for cryopreservation; and

(c) thawing the blood components, such that the stem cells are viable.

7. The composition of claim 6 which further comprises a human neonatal or fetal hematopoietic progenitor cell.

8. The composition of claim 6 in which the method of obtaining the cells further comprises the step after step (c) of removing a cryopreservative.

9. The composition of claim 6 in which the method of obtaining the cells further comprises the step after step (a) or step (c) of growing the stem cells in vitro.

10. The composition of claim 6 in which the method of obtaining the cells further comprises the step of enriching for stem and progenitor cells by a cell separation procedure.

11. The composition of claim 6 in which the blood components comprise whole neonatal or fetal blood.

12. The composition of claim 6 in which the blood components are isolated by collection from an umbilical cord.

13. The composition of claim 6 in which the blood components are isolated by collection from a placenta.

14. The composition of claim 6 in which the cryopreserving is by use of a cryopreservative.

15. The composition of claim 6 in which the cryopreservative is dimethyl sulfoxide.

16. The composition of claim 6 in which the cryopreserving is by use of liquid nitrogen.

17. A pharmaceutical composition comprising

(a) viable human neonatal or fetal hematopoietic stem cells derived from the umbilical cord blood or placental blood of a single human collected at the birth of said human, in which said cells are present in an amount sufficient to effect hematopoietic reconstitution of a human adult, which cells have been cryopreserved;

and

(b) a pharmaceutically acceptable carrier.

18. The composition of claim 17 which further comprises a viable human neonatal or fetal hematopoietic progenitor cell.

19. The composition of claim 17 which comprises whole neonatal or fetal blood.