Multi-lineage progenitor cells

• US 7,670,596 B2
• Filed: 04/20/2005
• Issued: 03/02/2010
• Est. Priority Date: 04/23/2004
• Status: Expired due to Fees

• Current Assignee: BioE LLC
• Original Assignee: BioE, Inc.
• Inventors: Sprague, Stacey L., Collins, Daniel P., Tigges, Barbara M.
• Primary Examiner(s): LEAVITT, MARIA GOMEZ
• Application Number:
  US11/110,299
• Publication Number:
  US 20050255592A1
• Time in Patent Office: 1,777 Days
• Field of Search: 424/93.1, 424/93.7, 424/3, 435/2, 435/7.23, 435/7.24, 435/7.25, 435/325, 435/355, 436/523
• US Class Current: 424/93.1
• CPC Class Codes: C12N 5/0607 Non-embryonic pluripotent stem cells, e.g. MASC induced pluripotent stem cells C12N5/0696

Abstract

Fetal blood multi-lineage progenitor cells that are capable of a wide spectrum of transdifferentiation are described.

Claims

1. A purified population of human fetal blood multi-lineage progenitor cells (MLPC), wherein said MLPC are positive for CD9, CD13, CD29, CD44, CD73, CD90 and CD105, and negative for CD10, CD34, CD41, CD45, Stro-1, Stage Specific Embryonic Antigen-3 (SSEA-3) and SSEA-4.

2. The purified population of MLPC of claim 1, wherein said MLPC display a fibroblast morphology.

3. The purified population of MLPC of claim 1, wherein said MLPC are obtained from cord blood.

4. The purified population of MLPC of claim 1, wherein said MLPC are further negative for CD2, CD3, CD4, CD5, CD7, CD8, CD14, CD15, CD16, CD19, CD20, CD22, CD33, CD36, CD38, CD61, CD62E, CD133, glycophorin-A, stem cell factor, and HLA-DR.

5. The purified population of MLPC of claim 1, wherein said MLPC adhere to a plastic surface when cultured.

6. The purified population of MLPC of claim 1, wherein said MLPC are capable of differentiating into cells from all three embryonic germ layers.

7. The purified population of MLPC of claim 6, wherein said MLPC are capable of differentiating into cells having an osteocytic phenotype, cells having an adipocytic phenotype, cells having a neural stem cell phenotype, cells having a myocytic phenotype, cells having an endothelial phenotype, cells having a hepatocyte precursor phenotype, and cells having a pancreatic precursor phenotype.

8. The purified population of MLPC of claim 1, wherein said MLPC comprise an exogenous nucleic acid.

9. The purified population of MLPC of claim 8, wherein said exogenous nucleic acid encodes a polypeptide.

10. A clonal line of human fetal blood MLPC, wherein said MLPC are positive for CD9, CD13, CD29, CD44, CD73, CD90 and CD105, and negative for CD10, CD34, CD41, CD45, Stro-1, SSEA-3 and SSEA-4.

11. The clonal line of claim 10, wherein said MLPC display a fibroblast morphology.

12. The clonal line of claim 10, wherein said MLPC are obtained from cord blood.

13. The clonal line of claim 10, wherein said MLPC are further negative for CD2, CD3, CD4, CD5, CD7, CD8, CD14, CD15, CD16, CD19, CD20, CD22, CD33, CD36, CD38, CD61, CD62E, CD133, glycophorin-A, stem cell factor, and HLA-DR.

14. The clonal line of claim 10, wherein said MLPC adhere to a plastic surface when cultured.

15. The clonal line of claim 10, wherein said MLPC are capable of differentiating into cells from all three embryonic germ layers.

16. The clonal line of claim 15, wherein said MLPC are capable of differentiating into cells having an osteocytic phenotype, cells having an adipocytic phenotype, cells having a neural stem cell phenotype, cells having a myocytic phenotype, cells having an endothelial phenotype, cells having a hepatocyte precursor phenotype, and cells having a pancreatic precursor phenotype.

17. The clonal line of claim 10, wherein said MLPC comprise an exogenous nucleic acid.

18. The clonal line of claim 17, wherein said exogenous nucleic acid encodes a polypeptide.

19. The clonal line of claim 10, wherein said MLPC have undergone at least 30 doublings in culture.

20. The clonal line of claim 10, wherein said MLPC have undergone at least 35 doublings in culture.

21. The clonal line of claim 10, wherein said MLPC have undergone at least 40 doublings in culture.

22. The clonal line of claim 10, wherein said MLPC have undergone at least 50 doublings in culture.

23. An article of manufacture comprising the purified population of MLPC of claim 14, or the clonal line of claim 10.

24. The article of manufacture of claim 23, wherein said purified population of MLPC or said clonal line is housed within a container.

25. The article of manufacture of claim 24, wherein said container is a vial.

26. The article of manufacture of claim 24, wherein said container is a bag.

27. A purified population of MLPC, wherein said MLPC are made by the following method:

a) contacting a human fetal blood sample with a composition, said composition comprising;

i) dextran;

ii) anti-glycophorin A antibody;

iii) anti-CD15 antibody; and

iv) anti-CD9 antibody;

b) allowing said sample to partition into an agglutinate and a supernatant phase;

c) recovering cells from said supernatant phase;

d) purifying MLPC from the recovered cells by adherence to a solid substrate; and

e) culturing said MLPC such that said MLPC obtain a fibroblast morphology, wherein said MLPC, after obtaining said fibroblast morphology, are positive for CD9, CD13, CD29, CD44, CD73, CD90 and CD105, and negative for CD10, CD34, CD41, CD45, Stro-1, SSEA-3, and SSEA-4.

28. The purified population of MLPC of claim 27, wherein said MLPC are obtained from cord blood.

29. The purified population of MLPC of claim 27, wherein said MLPC are further negative for CD2, CD3, CD4, CD5, CD7, CD8, CD14, CD15, CD16, CD19, CD20, CD22, CD33, CD36, CD38, CD61, CD62E, CD133, glycophorin-A, stem cell factor, and HLA-DR.

30. The purified population of MLPC of claim 27, wherein said MLPC are capable of differentiating into cells from all three embryonic germ layers.

31. The purified population of MLPC of claim 30, wherein said MLPC are capable of differentiating into cells having an osteocytic phenotype, cells having an adipocytic phenotype, cells having a neural stem cell phenotype, cells having a myocytic phenotype, cells having an endothelial phenotype, cells having a hepatocyte precursor phenotype, and cells having a pancreatic precursor phenotype.

32. A clonal line of MLPC, wherein said clonal line is made by the following method:

a) culturing a single MLPC in a culture vessel lacking other cells until a plurality of progeny are produced; and

b) culturing said progeny to obtain said clonal line of MLPC, wherein said MLPC are positive for CD9, CD13, CD29, CD44, CD73, CD90 and CD105, and negative for CD10, CD34, CD41, CD45, Stro-1, SSEA-3, and SSEA-4.

33. The clonal line of claim 32, wherein said MLPC are obtained from cord blood.

34. The clonal line of claim 32, wherein said MLPC are further negative for CD2, CD3, CD4, CD5, CD7, CD8, CD14, CD15, CD16, CD19, CD20, CD22, CD33, CD36, CD38, CD61, CD62E, CD133, glycophorin-A, stem cell factor, and HLA-DR.

35. The clonal line of claim 32, wherein said MLPC are capable of differentiating into cells from all three embryonic germ layers.

36. The clonal line of claim 35, wherein said MLPC are capable of differentiating into cells having an osteocytic phenotype, cells having an adipocytic phenotype, cells having a neural stem cell phenotype, cells having a myocytic phenotype, cells having an endothelial phenotype, cells having a hepatocyte precursor phenotype, and cells having a pancreatic precursor phenotype.