HUMAN MULTIPOTENT EMBRYONIC STEM CELL-LIKE PROGENITOR CELLS

• US 20120094380A1
• Filed: 10/18/2011
• Published: 04/19/2012
• Est. Priority Date: 10/18/2010
• Status: Active Grant

• Current Assignee: Sunshine Life Science & Technology Corp.
• Original Assignee: Sunshine Biotech Incorporated
• Inventors: TZU-BI SHIH, Daniel, Tsai, Ming-Song
• Granted Patent: US 9,234,177 B2
• Time in Patent Office: Days
• US Class Current: 435/366
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/999 Small molecules not provided for elsewhere
• 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
• CPC Class Codes: G01N 15/01 specially adapted for biological cells, e.g. blood cells investigating sedimentation of particle suspensions in blood G01N15/05
• CPC Class Codes: G01N 15/14 Optical investigation techniques, e.g. flow cytometry
• CPC Class Codes: G01N 15/149 specially adapted for sorting particles, e.g. by their size or optical properties

Abstract

The invention provides a plurality of embryonic stem cell-like progenitor cells, which are isolated from a human tissue by a systemic screening of human mesenchymal stromal stem/progenitor cells and a cell sorting by a cell antigen selected from the group consisting of CD34, CD117, CD133, CD201, GloboH and combination thereof, and cultured in a medium supplemented with at least one or more steroids and one or more growth factors. The cells of the invention express CD34 and exhibit sphere-like clonogenicity in early passages and express multipotent embryonic stem cells (ESCs) like characteristics.

Claims

1. An enriched population of multipotent human mesenchymal common progenitor cells (MCPCs), which are identified as mesenchymal stromal stem/progenitor cells, having at least the following characteristics:

CD14⁺, CD34⁺, CD117⁺, CD133⁺ (AC133⁺), CD201⁺, Nestin⁺, SSEA3⁺, SSEA4⁺ and GloboH⁺.

2. The enriched population of multipotent human MCPCs according to claim 1, wherein the MCPCs further have at least one of the following characteristics:

CD44⁺, CD54⁺, CD56⁺, CD105⁺, CD146⁺, and PDGFR⁺.

3. The enriched population of multipotent human MCPCs according to claim 1, which exhibit sphere-like clonogenicity in early passages and express multipotent embryonic stem cells (ESCs) like characteristics.

4. The enriched population of multipotent human MCPCs according to claim 1, which are mesenchymal stromal stem/progenitor cells having the following characteristics:

CD14⁺, CD34⁺, CD117⁺, CD133⁺ (AC133⁺), CD201⁺, Nestin⁺, SSEA3⁺, SSEA4⁺, GloboH⁺, CD44⁺, CD54⁺, CD56⁺, CD105⁺, CD146⁺ and PDGFR⁺.

5. The enriched population of multipotent human MCPCs according to claim 1, which is isolated from a human somatic tissue.

6. The enriched population of multipotent human MCPCs according to claim 5, wherein the human somatic tissue is selected from the group consist of amnion, chorion, umbilical cord, endometrium, gingival, bone marrow, and adipose.

7. The enriched population of multipotent human MCPCs according to claim 6, where the human somatic tissue is amnion.

8. The enriched population of multipotent human MCPCs according to claim 1, which has the potentials to differentiate into ectodermal cells, mesodermal cells, and endodermal cells.

9. The enriched population of multipotent human MCPCs according to claim 1, which has the potentials of adipogenic differentiation, osteogenic differentiation, chondrogenic differentiation, neurogenic differentiation, cardiomyogenic differentiation, endothelial differentiation, and hepatic differentiation.

10. A method for producing the enriched population of multipotent human MCPCs according to claim 1, comprising:

isolating from a human somatic tissue by a systemic screening of human mesenchymal stromal stem/progenitor cells followed by a cell sorting by a cell antigen selected from the group consisting of CD34, CD117, CD133, CD201, GloboH and combination thereof; and

culturing in a medium supplemented with at least one or more steroids and one or more growth factors.

11. The method according to claim 10, wherein the human somatic tissue is selected from the group consist of amnion, chorion, umbilical cord, endometrium, gingival, bone marrow, and adipose

12. The method according to claim 10, wherein the cell sorting is a fluorescence-activated cell sorting (FACS) flow cytometry.

13. The method according to claim 10, wherein the steroid is one selected from the group consisting of a corticosteroid, cholesterol and combination thereof.

14. The method according to claim 10, wherein the growth factor is one selected from the group consisting of epidermal growth factor (EGF), fibroblast growth factor (FGF), insulin-like growth factor (IGF), insulin, platelet-derived growth factor (PDGF), IL-6, thrombopoietin (TPO), and combination thereof.

15. A composition comprising the enriched population of multipotent human MCPCs according to claim 1, which are encapsulated in alginate.

16. A feeder cell layer for stem cell culture comprising the enriched population of multipotent human MCPCs according to claim 1.

17. A stem cell niche comprising the enriched population of multipotent human MCPCs according to claim 1 seeded on a scaffold.