Multipotent Prenatal Stem Cells

• US 20150216911A1
• Filed: 09/26/2013
• Published: 08/06/2015
• Est. Priority Date: 09/26/2013
• Status: Active Grant

• Current Assignee: Prime Merger Sub LLC (Alight, Inc.)
• Original Assignee: NuTech Spine, Inc.
• Inventors: Vines, Jeremy B., Walthall, Howard P. Jr.
• Granted Patent: US 9,700,585 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 35/50 Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
• CPC Class Codes: C12N 5/0605 Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly
• CPC Class Codes: C12N 5/0668 Mesenchymal stem cells from other natural sources

Abstract

The present invention relates to the isolation, propagation and use of prenatal stem cells. The cells are CD105 negative. In certain embodiments the cells are also SSEA-4 positive or negative, and c-kit negative. The cells may be isolated from a prenatal sample, for instance third trimester amniotic fluid or placental membrane. The cells may be combined with a biologically compatible solution or a biologically compatible matrix and utilized to treat a human subject. The invention also contemplated enriched populations of prenatal stem cells. The enriched cell population may be propagated in the appropriate culture medium and the resulting progeny cells utilized therapeutic applications.

Claims

1. An isolated human prenatal stem cell that is CD105 negative.

2. The isolated human prenatal stem cell of claim 1, wherein the cell is c-kit negative.

3. The isolated human prenatal stem cell of claim 1, wherein the cell is isolated from a prenatal sample.

4. The isolated human prenatal stem cell of claim 3, wherein the prenatal sample comprises prenatal fluid.

5. The isolated human prenatal stem cell of claim 4, wherein the prenatal fluid comprises third trimester amniotic fluid.

6. The isolated human prenatal stem cell of claim 3, wherein the prenatal sample comprises prenatal membrane.

7. The isolated human prenatal stem cell of claim 6, wherein the prenatal membrane comprises placental membrane.

8. The isolated human prenatal stem cell of claim 1, wherein the cell is SSEA-4 negative.

9. The isolated human prenatal stem cell of claim 1, wherein the cell is SSEA-positive.

10. A preparation comprising the isolated human prenatal stem cell of claim 1 and a biologically compatible solution.

11. A preparation comprising the isolated human prenatal stem cell of claim 1 imbedded into an implantable biologically compatible matrix.

12. A method of treating a human subject comprising injecting the preparation of claim 10 into the bloodstream of the human subject.

13. A method of treating a human subject comprising implanting the preparation of claim 11 into the human subject.

14. An enriched population of isolated human prenatal stem cells wherein the population is essentially CD105 negative.

15. The enriched population of claim 14, wherein the population is essentially c-kit negative.

16. The enriched population of claim 14, wherein the cells are isolated from a third-trimester amniotic fluid sample.

17. The enriched population of claim 14, wherein the cells are isolated from a placental membrane.

18. The enriched population of claim 14, wherein the population is essentially SSEA-4 negative.

19. The enriched population of claim 14, wherein the population is essentially SSEA-4 positive.

20. A preparation comprising the enriched population of claim 14 and a biologically compatible solution.

21. A preparation comprising the enriched population of claim 14 imbedded into an implantable biologically compatible matrix.

22. A method of treating a human subject comprising injecting the preparation of claim 20 into the bloodstream of the subject.

23. A method of treating a human subject comprising implanting the preparation of claim 21 into the subject.

24. A method of proliferating a population of cells enriched for human prenatal stem cells comprising:

selecting at least one cell from a human prenatal sample, wherein the cell is CD105 negative;

introducing the at least one cell to a culture medium; and

proliferating the at least one cell in the culture medium.

25. The method of claim 24, wherein selecting the at least one cell comprises the steps of:

incubating a plurality of cells from the human prenatal sample with a first antibody that is reactive to CD105; and

selecting the at least one cell from the plurality of cells.

26. The method of claim 25, further comprising the steps of:

incubating the at least one cell with a second antibody that is reactive to SSEA-4 thereby producing a group of cells; and

selecting from the group of cells a number of cells that are non-reactive with the second antibody.

27. The method of claim 25, further comprising the steps of:

incubating the at least one cell with a second antibody that is reactive to SSEA-4 thereby producing a group of cells; and

selecting from the group of cells a number of cells that are reactive with the second antibody.

28. The method of claim 25, wherein selecting the at least one cell from the plurality of cells is accomplished by an immunoselection technique.

29. The method of claim 28, where the immunoselection technique is selected from the group consisting of flow cytometry, activated cells sorting and immune-magnetic selection.

30. The method of claim 25, wherein the at least one cell is c-kit negative.

31. The method of claim 25, further comprising isolating the at least one cell from a third-trimester human amniotic fluid sample.

32. The method of claim 25, further comprising isolating the at least one cell from a prenatal membrane.

33. A method of harvesting human prenatal stem cells comprising:

providing a human prenatal sample;

collecting cells from the sample; and

selecting CD105 negative cells from the sample.

34. The method of claim 33, wherein the prenatal sample comprises prenatal fluid.

35. The method of claim 34, wherein the prenatal fluid comprises third trimester amniotic fluid.

36. The method of claim 33, wherein the prenatal sample comprises prenatal membrane.

37. The method of claim 36, wherein the prenatal membrane comprises placental membrane.

38. The method of claim 33, wherein the selecting of the CD105 negative cells comprises the steps of:

incubating a plurality of cells from the human prenatal sample with a first antibody reactive to SSEA-4; and

selecting from the plurality of cells a group of cells that are reactive with the first antibody.

39. The method of claim 33, further comprising the steps of:

incubating the group of cells that are reactive with the first antibody with a second antibody that is reactive to CD105; and

selecting from the group of cells that are reactive with the first antibody a second group of cell that are non-reactive with the second antibody.

40. The method of claim 38, wherein the selecting from the plurality of cells a group of cells that are reactive with the first antibody is accomplished by an immunoselection technique.

41. The method of claim 40, where the immunoselection technique is selected from the group consisting of flow cytometry, activated cells sorting and immune-magnetic selection.

42. The method of claim 33, further comprising selecting c-kit negative cells from the CD105 negative cells.