Growth of embryonic stem cells

• US 20080003676A1
• Filed: 06/11/2007
• Published: 01/03/2008
• Est. Priority Date: 06/26/2006
• Status: Abandoned Application

• Current Assignee: Millipore Corporation (Merck & Co., Inc.)
• Original Assignee: Millipore Corporation (Merck & Co., Inc.)
• Inventors: Sheridan, Steven, Gil, Sonia
• Application Number:
  US11/811,532
• Publication Number:
  US 20080003676A1
• Time in Patent Office: Days
• US Class Current: 435/354
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 39/00 General protective or antinoxious agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 2502/02 embryonic cells
• CPC Class Codes: C12N 2502/13 connective tissue cells; generic mesenchyme cells, e.g. so-called "embryonic fibroblasts"
• CPC Class Codes: C12N 2533/30 Synthetic polymers
• CPC Class Codes: C12N 5/0606 Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696

Abstract

The invention provides systems, kits and methods relating to the growth of stem cells.

Claims

1. A method of growing an embryonic stem cells (ESC) in vitro comprising a) contacting a surface of a porous membrane with an ESC;

b) contacting the surface of a solid support with a feeder cell;

c) providing a culture media to the ESC;

d) providing a culture media to the feeder cell, wherein the porous membrane is in fluid communication with the culture media of d).

2. The method of claim 1, wherein the ESCs are maintained in an undifferentiated state.

3. The method of claim 1, wherein the ESCs are chosen from murine embryonic stem cells and human embryonic stem cells.

4. The method of claim 1, wherein the feeder cell is a fibroblast.

5. The method of claim 4, wherein the fibroblast is chosen from a murine and a human fibroblast.

6. The method of claim 5, wherein the fibroblast is an embryonic fibroblast.

7. The method of claim 1, wherein the ESC culture media comprises serum.

8. The method of claim 1, wherein the ESC culture media optionally comprises LIF.

9. The method of claim 1, wherein in the porous membrane and the solid support are provided in a multiwell plate.

10. The method of claim 9, wherein the multiwell plate is chosen from a plate comprised of 6, 12, 48 or 96 wells.

11. The method of claim 1, wherein the porous membrane is comprised of a polymer.

12. The method of claim 11, wherein the polymer is chosen from polyethylene terephthalate, polymer cladded fibers, polyether sulfone, polyamide, agarose, cellulose, a polysaccharide, polytetrafluoroethylene, polysulfone, polyester, polyvinylidene fluoride, polypropylene, a fluorocarbon, e.g. poly(tetrafluoroethylene-co-perfluoro(alkyl vinyl ether)), poly carbonate, polyethylene, glass, ceramic, nylon and metal.

13. A system for growing ESCs in vitro comprising a solid support;

a porous membrane in fluid communication with the solid support; and

a culture media suitable for growing ESCs.

14. The system of claim 13 further comprising embryonic fibroblasts.

15. The system of claim 13, wherein in the porous membrane and the solid support are provided in a multiwell plate.

16. The system of claim 15, wherein the multiwell plate is chosen from a plate comprised of 6, 12, 48 or 96 wells.

17. The system of claim 13, further comprising ESCs.

18. The system of claim 17, wherein the ESCs are chosen from murine ESCs and human ESCs.

19. A kit for growing ESCs comprising a solid support;

a porous membrane in fluid communication with the solid support;

at least one container; and

instructions for growing ESCs.

20. The kit of claim 19, wherein in the porous membrane and the solid support are provided in a multiwell plate.

21. A method of growing an ESC in a tissue culture plate comprising one or more wells, where each of the wells comprises a porous membrane insert and a solid support positioned in fluid communication with the porous membrane insert, and where the method comprises a) contacting a surface of the porous membrane with an ESC;

b) contacting a surface of the solid support, which lies beneath the porous membrane insert with an embryonic fibroblast;

c) providing a culture media to the ESC;

d) providing a culture media to the embryonic fibroblast.

22. The method of claim 1, wherein the media is serum free.