Progenitor cells from wharton's jelly of human umbilical cord

• US 20050148074A1
• Filed: 10/08/2004
• Published: 07/07/2005
• Est. Priority Date: 02/11/2003
• Status: Active Grant

• Current Assignee: Tissue Regeneration Therapeutics Inc
• Original Assignee: Tissue Regeneration Therapeutics Inc
• Inventors: Davies, John E., Baksh, Dolores, Lickorish, Antony D.S., Sarugaser, Rahul, Hosseini, Morris
• Granted Patent: US 7,547,546 B2
• Time in Patent Office: Days
• US Class Current: 435/372
• CPC Class Codes: A61K 35/51 Umbilical cord; Umbilical cord blood; Umbilical stem cells
• CPC Class Codes: C12N 2509/00 Methods for the dissociation of cells, e.g. specific use of enzymes
• CPC Class Codes: C12N 2523/00 Culture process characterised by temperature
• CPC Class Codes: C12N 5/0605 Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly
• CPC Class Codes: C12N 5/0653 Adipocytes; Adipose tissue
• CPC Class Codes: C12N 5/0654 Osteocytes, Osteoblasts, Odontocytes; Bones, Teeth
• CPC Class Codes: C12N 5/0668 Mesenchymal stem cells from other natural sources

Abstract

Human progenitor cells are extracted from perivascular tissue of human umbilical cord. The progenitor cell population proliferates rapidly, and harbours osteogenic progenitor cells and MHC−/− progenitor cells, and is useful to grow and repair human tissues including bone.

Claims

1. A Wharton'"'"'s jelly extract, wherein the extract comprises human progenitor cells and is obtained by enzymatic digestion of the perivascular tissue proximal to the vasculature of human umbilical cord.

2. A Wharton'"'"'s jelly extract according to claim 1, wherein the extract is essentially free from cells of umbilical cord blood.

3. A Wharton'"'"'s jelly extract according to claim 2, wherein the extract is obtained by subjecting umbilical cord vasculature bearing proximal Wharton'"'"'s jelly to enzymatic digestion in a suitable cell extraction medium.

4. A Wharton'"'"'s jelly extract according to claim 3, wherein the step of enzymatic digestion results in the release of cells from the collagen matrix of the Wharton'"'"'s jelly.

5. A Wharton'"'"'s jelly extract according to claim 4, in which the extraction is performed at 37°

C. in phosphate buffered saline and 0.5-10.0 mg/mL collagenase for a period of 1 to 24 hours.

6. A method for obtaining a human progenitor cell, comprising the step of isolating said cell from the Wharton'"'"'s extract according to claim 1.

7. A method for producing a cell population comprising human progenitor cells, the method comprising the step of culturing an isolated human progenitor cell obtained by the method according to claim 6.

8. A cell population comprising human progenitor cells, whenever obtained by the method according to claim 7.

10. A cell population enriched for osteoprogenitor cells obtained by the method according to claim 7.

11. A cell population enriched for immuno-incompetent human progenitor cells obtained by the method according to claim 7.

9. A cell population useful as a source of human progenitor cells, wherein the cell population is extractable from perivascular tissue of human umbilical cord, and has the characteristics of (1) rapid proliferation, (2) the presence of osteoprogenitor cells, and (3) the presence of immuno-incompetent cells.

12. A method for producing bone tissue, comprising the step of subjecting a cell population according to claim 9 to culturing in the absence of osteogenic supplements.

13. A method for producing a population of MHC double negative human progenitor cells, comprising the step of culturing a previously frozen cell population defined in claim 9.

14. An isolated population of human progenitor cells, in the form of human umbilical cord perivascular cells having a 3G5+, CD45−

phenotype, the population comprising cells characterized by rapid proliferation, the presence of human osteoprogenitor cells, and the presence of human progenitor cells that are negative for markers of MHC class I and MHC class II.

15. An isolated population of human progenitor cells, the population comprising cells that upon culturing give rise spontaneously to cells of a type selected from bone cells and fat cells, the population comprising the non-adherent fraction of the cell population according to claim 14.

19. A composition comprising an effective amount of a cell population according to claim 14, and a carrier suitable for delivering said population to a tissue site.

20. A method of tissue engineering, comprising the step of delivering to a tissue site at which engineering is desired, a composition according to claim 19.

16. An isolated population of human progenitor cells, wherein at least 50% of the cells in said population have an MHC double negative phenotype.

17. An isolated population of human progenitor cells according to claim 16, wherein at least 80% of the cells in said population have an MHC double negative phenotype.

18. An isolated population of human progenitor cells according to claim 17, wherein at least 90% of the cells in said population have an MHC double negative phenotype.