Isolation and preservation of fetal hematopoietic and mesencymal system cells from non-controversial materials and/or tissues resulting from miscarriages and methods of therapeutic use

• US 20080171019A1
• Filed: 01/17/2007
• Published: 07/17/2008
• Est. Priority Date: 01/17/2007
• Status: Active Grant

• Current Assignee: Maria Michejda
• Original Assignee: Maria Michejda
• Inventors: Michejda, Maria
• Granted Patent: US 7,883,698 B2
• Time in Patent Office: Days
• US Class Current: 424/93.7
• CPC Class Codes: A01N 1/0221 Freeze-process protecting agents, i.e. substances protecting cells from effects of the physical process, e.g. cryoprotectants, osmolarity regulators like oncotic agents
• CPC Class Codes: A61K 35/48 Reproductive organs

Abstract

The present invention is directed to compositions of fetal hematopoietic stem and progenitor cells and stromal (mesencymal) cells derived from second trimester (16-20 weeks gestation) miscarriages, that are isolated, processed and cryopreserved, and the therapeutic uses of such stem and progenitor cells upon application. Such compositions may be useful for hematopoietic reconstitution and/or replacement in patients with various diseases and metabolic disorders. The invention also relates to methods for collecting, isolating, processing and cryopreserving of the fetal stem and progenitor cells of the invention. Importantly, the invention meets an urgent need in regenerative medicine while at the same time ensures utility of the product compositions because it utilizes fetal materials from second trimester (16-20 weeks of gestation) spontaneously lost pregnancies which are free of moral or ethical burdens.

Claims

1. A method for collection of human fetal tissue from second trimester miscarriages (16-20 weeks of gestation), comprising:

(a) collection of human fetal bone marrow that contains hematopoietic stem or progenitor cells and stromal (mesencymal) cells;

(b) collection of other fetal tissues for therapies of life threatening diseases;

(c) cryopreservation of hematopoietic or progenitor cells, as well as other fetal tissues, utilizing a non-toxic cryopreservation cocktail consisting of 2% or less dimethyl sulfoxide, human serum albumin and dextran in RPMI medium; and

(d) thawing of the cryopreserved hematopoietic or progenitor cells, such that the stem or progenitor cells are at least 90% viable.

2. The method according to claim 1, further comprising testing for possible genetic, viral, fungal and bacterial contamination of fetal tissue components.

3. The method according to claim 1 further comprising the step of mixing the hematopoietic or progenitor cells with fetal bone chips in an appropriate matrix to produce a bone paste.

6. The method according to claim 1 in which the stem cells are autologous to the host.

7. The method according to claim 1 in which the stem cells are syngeneic to the host.

8. The method according to claim 1 in which the stem cells are allogeneic to the host.

9. The method according to claim 1 in which the a host has a disease selected from the group consisting of immunodeficiencies, a malignant solid tumors, anemias, including but not limited to sickle cell anemia, aplastic anemia and Fanconi'"'"'s anemia, hyperproliferative stem cell disorder, hematopoietic malignancies such as leukemias, lymphomas, autoimmune disease, hemolytic disorder, genetic disorders such as thalassemia.

10. The method according to claim 1 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.

4. (canceled)

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

(a) isolating human fetal bone marrow stem cells from 16-20 weeks miscarriages; and

(b) introducing the fetal bone marrow cells into suitable human host, such that the fetal stem cells are viable and can proliferate within the host.

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

(a) isolating human fetal components from 16-20 week miscarriages, containing hematopoietic stem and progenitor cells;

(b) cryopreserving the hematopoietic components, utilizing a non-toxic cryopreservation cocktail consisting of 2% or less dimethyl sulfoxide, human serum albumin and dextran in RPMI medium;

(c) thawing the hematopoietic components; and

(d) introducing the fetal cells into a suitable human host, such that the hematopoietic stem and progenitor cells are viable and can proliferate and reconstitute within the host.

12. The method according to claim 11 in which the stem and progenitor cells are autologous to the host.

13. The method according to claim 12 in which the stem and progenitor cells are syngeneic to the host.

14. The method according to claim 12 in which the stem and progenitor cells are allogeneic to the host.

15. A method for hematopoietic or immune reconstitution of a human comprising introducing into the human a composition comprising human fetal hematopoietic stem cells derived from fetal components from 16-20 week miscarriages, in which the stem cells are progeny of cells which have been previously cryopreserved , utilizing a non-toxic cryopreservation cocktail consisting of 2% or less dimethyl sulfoxide, human serum albumin and dextran in RPMI medium.