rbLIF PROTEIN FOR USE IN EMBRYONIC STEM CELL CULTURES

• US 20100227403A1
• Filed: 12/14/2009
• Published: 09/09/2010
• Est. Priority Date: 12/15/2008
• Status: Abandoned Application

• Current Assignee: Renova Life, Inc.
• Original Assignee: Renova Life, Inc.
• Inventors: Yang, Jerry, Xu, Jie, Tian, Xiuchun, Ma, Yinghong, Du, Fuliang
• Application Number:
  US12/637,660
• Publication Number:
  US 20100227403A1
• Time in Patent Office: Days
• US Class Current: 435/375
• CPC Class Codes: C07K 14/5415 Leukaemia inhibitory factor [LIF]
• CPC Class Codes: C12N 2501/235 Leukemia inhibitory factor [LIF]

Abstract

Provided herein methods of maintaining an embryonic stem cell culture in an undifferentiated state, wherein the method includes culturing an embryonic stem cell on a rabbit feeder cell layer, wherein the rabbit feeder layer expresses a leukemia inhibitory factor (LIF) protein. Also provided are methods of maintaining an embryonic stem cell culture in an undifferentiated state, wherein the method includes culturing an embryonic stem cell in a culture medium comprising rabbit leukemia inhibitory factor (rbLIF) protein. Also provided are polynucleotide and amino acid sequences of a rabbit leukemia inhibitory factor protein (rbLIF).

Claims

1. A rabbit leukemia inhibitory factor (rbLIF) protein comprising the amino acid sequence set forth in SEQ ID NO:

5.

2. A polynucleotide encoding the protein of claim 1.

3. The polynucleotide of claim 2, wherein the polynucleotide comprises the sequence set forth in SEQ ID NO:

4.

4. A method of maintaining an embryonic stem cell culture in an undifferentiated state, comprising culturing an embryonic stem cell on a rabbit feeder cell layer, wherein the rabbit feeder layer expresses an leukemia inhibitory factor (LIF) protein.

5. The method of claim 4, wherein the culturing is in a culture medium supplemented with LIF protein.

6. The method of claim 4, wherein the feeder cell layer expresses a selection marker.

7. The method of claim 6, wherein the selection marker is an antibiotic resistance gene.

8. The method of claim 4, wherein the LIF protein is rbLIF.

9. The method of claim 4, wherein the embryonic stem cells are from rabbit.

10. The method of claim 4, wherein the embryonic stem cells are from mouse.

11. The method of claim 4, wherein the rabbit feeder cell layer is obtained from embryonic tissues from rabbit embryos of 3 weeks of development or less.

12. The method of claim 11, wherein the rabbit embryos are obtained from a transgenic rabbit expressing a transgene encoding a selection marker.

13. The method of claim 12, wherein the selection marker is an antibiotic resistance gene.

14. The method of claim 13, wherein the antibiotic resistance gene is a neomycin resistance gene.

15. A method of maintaining an embryonic stem cell culture in an undifferentiated state, comprising culturing an embryonic stem cell in a culture medium comprising rabbit leukemia inhibitory factor (rbLIF) protein.

16. The method of claim 15, wherein the culture medium is conditioned medium comprising rbLIF protein.

17. The method of claim 15, wherein the embryonic stem cells are from rabbit.

18. The method of claim 15, wherein the embryonic stem cells are from mouse.