Early stage multipotential stem cells in colonies of bone marrow stromal cells

• US 7,056,738 B2
• Filed: 03/23/2001
• Issued: 06/06/2006
• Est. Priority Date: 03/23/2001
• Status: Expired due to Fees

• Current Assignee: Drexel University, Tulane University
• Original Assignee: Philadelphia Health and Education Corporation, Tulane University
• Inventors: Colter, David C., Sekiya, Ichiro, Prockop, Darwin J.
• Primary Examiner(s): FALK, ANNE MARIE
• Application Number:
  US09/816,182
• Publication Number:
  US 20020168765A1
• Time in Patent Office: 1,901 Days
• Field of Search: 435/325, 435/366, 435/372, 435/377, 435/353, 435/354
• US Class Current: 435/372
• CPC Class Codes: C12N 5/0663 Bone marrow mesenchymal stem cells (BM-MSC)
• CPC Class Codes: C12Q 1/6881 for tissue or cell typing, e.g. human leukocyte antigen [HLA] probes

Abstract

Marrow stromal cells (MSCS) are adult stem cells from bone marrow that can differentiate into multiple non-hematopoietic cell lineages. Colonies of human MSCs were shown to contain both small, rapidly self-renewing stem cells (RS cells) and large, more mature cells (mMSCs). Samples enriched for RS cells had a greater potential for multipotential differentiation than samples enriched for mMSCs. Also, RS cells have a series of surface epitopes and expressed proteins that can be used to differentiate RS cells from mMSCs. The results suggest that it will be important to distinguish the two major sub-populations of MSCs in defining their biology and their potentials for cell and gene therapy.

Claims

1. A population of human marrow stromal cells enriched for small and rapidly self-renewing stem (RS) cells, said population comprising about 95% RS cells, wherein said RS cells are about seven microns in diameter, comprise granular and agranular cells, and do not express STRO-1, PDGF-R, EGF-R, CD10 and CD147.

2. The population of human marrow stromal cells of claim 1, wherein said population enriched for RS cells has an increased capacity for multilineage differentiation compared to a population of human marrow stromal cells not enriched for RS cells.

3. The population of human marrow stromal cells of claim 1, wherein said RS cells have a high nucleus to cytoplasm ratio.

4. The population of human marrow stromal cells of claim 1, further wherein said RS cells express at least one of the polypeptides selected from the group consisting of VEGF receptor-2 (FLK-1), TRK (an NGF receptor), transferrmn receptor, annexin II (lipocortin 2) and CD49e (integrin alpha 5).

5. The population of human marrow stromal cells of claim 1, further wherein cells within a first subtype of said RS cells are agranular and express VEGF receptor-2 (FLK-1), TRK (an NGF receptor), transferrin receptor, and annexin II.

6. The population of human marrow stromal cells of claim 5, further wherein said cells within a first subtype express CD44 (Hyaluronic acid receptor), CD49e (integrin alpha 5) and CD59.

7. The population of human marrow stromal cells of claim 1, further wherein cells within a second subtype of said RS cells are granular, express CD49e (integrin alpha 5) and do not express CD44 (Hyaluronic acid receptor) and CD59.

8. The population of human marrow stromal cells of claim 7, further wherein said cells within a second subtype do not express CD81 and CD90.

9. The population of human marrow stromal cells of claim 1, further wherein said RS cells express heat shock protein-27, tumor rejection antigen, glutathione-S transferase, peroxiredoxin 1, voltage-dependent-anion channel-2, protein kinase C substrate, phosphatase 2A inhibitor, esterase D, RNase A, initiation factor 5a, elongation factor 1-alpha, ribosomal protein S12, ribosomal protein large P1, ribosomal protein large P2, transcription factor BTF 3a, annexin I, destrin, myosin light chain, lactate dehydrogenase A, glycerolaldehyde-3-P dehydrogenase, citrate synthetase, transketolase, P-glycerolmutase, aldo-keto reductase 7(A2), alpha-amylase inhibitor CM3, enoyl-CoA hydratase, and proteosome subunit alpha-4.