Postpartum cells derived from placental tissue, and methods of making and using the same

Patent metadata

Field	Value
Publication number	US-10195233-B2
Application number	US-201715443718-A
Country	US
Kind code	B2
Filing date	Feb 27, 2017
Priority date	Jun 27, 2003
Publication date	Feb 5, 2019
Grant date	Feb 5, 2019

Abstract

Cells derived from postpartum placenta and methods for their isolation are provided by the invention. The invention further provides cultures and compositions of the placenta-derived cells. The placenta-derived cells of the invention have a plethora of uses, including but not limited to research, diagnostic, and therapeutic applications.

First claim

What is claimed:

1. A method of isolating a placenta-derived cell from a human postpartum placenta comprising dissociating said cell from said placenta with a matrix metalloprotease, a neutral protease, and a mucolytic enzyme that digests hyaluronic acid, wherein the isolated placenta-derived cell is obtained from human postpartum placenta substantially free of blood, and wherein the placenta-derived cell has the following characteristics:
a) self-renews and expands in culture;
b) is multipotent;
c) produces CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, PD-L2 and HLA-A,B,C;
d) does not produce CD31, CD34, CD45, CD80, CD86, CD117, CD141, CD178, B7-H2, HLA-G or HLA-DR-DP,DQ; and
e) expresses, relative to a human fibroblast, mesenchymal stem cell, or iliac crest bone marrow cell, increased levels of oxidized low density lipoprotein receptor 1 and renin.

2. The method of claim 1 , wherein said matrix metalloprotease comprises collagenase.

3. The method of claim 1 , wherein said neutral protease comprises dispase.

4. The method of claim 1 , wherein said mucolytic enzyme comprises hyaluronidase.

5. The method of claim 1 , further comprising mechanically dissociating said placenta prior to said dissociating step.

6. The method of claim 1 , further comprising growing said cell in culture medium.

7. The method of claim 6 , wherein said culture medium comprises RPMI1640, Ham's F10 medium, Ham's F12 medium, Mesenchymal Stem Cell Growth Medium, Iscove's modified Dulbecco's medium, Dulbecco's modified Eagle's Medium (DMEM), CELL-GRO FREE, DMEM/F12, advanced DMEM, DMEM/MCDB201, or Eagle's basal medium.

8. The method of claim 6 , wherein said medium comprises about 2% to about 15% (v/v) serum.

9. The method of claim 6 , wherein said medium comprises beta-mercaptoethanol.

10. The method of claim 6 , wherein said culture medium comprises at least one antibiotic agent.

11. The method of claim 6 , wherein said culture medium comprises glucose.

12. The method of claim 6 , wherein said culture medium comprises L-valine.

13. The method of claim 6 , wherein said culture medium comprises DMEM-low glucose, beta-mercaptoethanol, serum, and an antibiotic agent.

14. The method of claim 1 , further comprising cryopreserving said placenta-derived cell.

15. The method of claim 1 , further comprising banking said placenta-derived cell.

16. The method of claim 1 , further comprising expanding said isolated cells on an uncoated surface.

17. The method of claim 1 , further comprising expanding said isolated cells on a coated surface.

18. The method of claim 17 , wherein said surface is coated with at least one of gelatin, collagen, fibronectin, laminin, ornithine, vitronectin, or extracellular membrane protein.

19. The method of claim 6 , wherein the culture medium is protein-free culture medium.

20. The method of claim 6 , wherein the culture medium is serum-free culture medium.

21. The method of claim 1 , wherein the placenta-derived cell lacks production of GRO-alpha and oxidized low density lipoprotein receptor, as detected by flow cytometry.

22. The method of claim 1 , wherein the placenta-derived cell further has the following characteristics:
(a) secretion of monocyte chemotactic protein 1 (MCP-1), interleukin-6 (IL-6), interleukin 8 (IL8), hepatocyte growth factor (HGF), keratinocyte growth factor (KGF), heparin-binding epidermal growth factor (HB-EGF), brain-derived neurotrophic factor (BDNF), tissue inhibitor of matrix metalloproteinase 1 (TIMP1), thrombopoietin (TPO), macrophage inflammatory protein 1alpha(MIP1a), Rantes (regulated on activation, normal T cell expressed and secreted), thymus and activation-regulated chemokine (TARC), and Eotaxin; and
(b) lack of secretion of fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF), angiopoietin 2 (ANG2), platelet derived growth factor (PDGF-bb), transforming growth factor beta2 (TGFbeta2), macrophage inflammatory protein 1beta (MIP1b), I309, and macrophage-derived chemokine (MDC), as detected by ELISA.

23. The method of claim 1 , wherein the placenta-derived cell is of a neonatal origin.

24. The method of claim 1 , wherein said placenta-derived cell is of a maternal origin.

25. The method of claim 1 , wherein the placenta-derived cell has the ability to differentiate into a mesodermal, ectodermal, or endodermal phenotype.

26. The method of claim 1 , wherein the placenta-derived cell can undergo at least 40 population doublings in culture.

27. The method claim 1 , further comprising genetically engineering the isolated cell to produce a protein of interest.

28. The method of claim 1 , wherein the isolated placenta-derived cell is identified by ATCC Accession No. PTA-6074.

29. The method of claim 1 , wherein the isolated placenta-derived cell is identified by ATCC Accession No. PTA-6075.

30. The method of claim 1 , wherein the isolated placenta-derived cell is identified by ATCC Accession No. PTA-6079.

31. The method of claim 1 , wherein the isolated placenta-derived cell expresses granulocyte chemotactic protein-2 (GCP-2).

Assignees

• Depuy Synthes Products Inc

Inventors

• Kihm Anthony J
• Harris Ian Ross
• Mistry Sanjay
• Harmon Alexander M
• Messina Darin J
• Seyda Agnieszka
• Yi Chin-Feng
• Gosiewska Anna

Classifications

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P39/06

  Free radical scavengers or antioxidants · CPC title

• A61P37/06

  Immunosuppressants, e.g. drugs for graft rejection · CPC title

• A61P37/02

  Immunomodulators · CPC title

• A61P9/10

  for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis · CPC title