US2010041137A1PendingUtilityA1

Culture medium containing kinase inhibitors, and uses thereof

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Assignee: UNIV EDINGBURGHPriority: Mar 30, 2006Filed: Oct 20, 2009Published: Feb 18, 2010
Est. expiryMar 30, 2026(expired)· nominal 20-yr term from priority
Inventors:Austin Gerard SmithQi-Long Ying
C12N 2500/90C12N 2501/727C12N 5/0606C12N 2501/115C12N 5/0031
59
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Claims

Abstract

Pluripotent cells are maintained in a self-renewing state in serum-free culture medium comprising a MEK inhibitor, a GSK3 inhibitor and, optionally, an antagonist of an FGF receptor. Pluripotent cells are also maintained in a self-renewing state in serum-free culture medium comprising a MEK inhibitor and an antagonist of an FGF receptor.

Claims

exact text as granted — not AI-modified
1 . A method of culturing mouse or human pluripotent cells, comprising culturing the mouse or human pluripotent cells in the presence of (i) a MEK1 inhibitor, and (ii) CHIR99021, wherein the mouse or human pluripotent cells undergo self-renewal. 
   
   
       2 . The method of  claim 1 , wherein the MEK1 inhibitor is selected from PD184352 and PD98059. 
   
   
       3 . A method of obtaining pluripotent cells, comprising culturing cells in a culture medium comprising a MEK inhibitor and a GSK3 inhibitor, thereby obtaining pluripotent cells. 
   
   
       4 . The method of  claim 3 , wherein the culture medium further comprises an antagonist of an FGF receptor. 
   
   
       5 . The method of  claim 3 , wherein the cells comprise mouse cells. 
   
   
       6 . The method of  claim 3 , wherein the cells comprise human cells. 
   
   
       7 . The method of  claim 3 , wherein the cells comprise rat cells. 
   
   
       8 . The method of  claim 3 , comprising culturing the cells in the absence of serum. 
   
   
       9 . The method of  claim 3 , wherein the MEK inhibitor is a MEK1 inhibitor. 
   
   
       10 . The method of  claim 9 , wherein the MEK inhibitor is selected from PD184352 and PD98059. 
   
   
       11 . The method of  claim 3 , wherein the GSK3 inhibitor is an inhibitor of GSK-3β. 
   
   
       12 . The method of  claim 11 , wherein the GSK3 inhibitor is CHIR99021. 
   
   
       13 . The method of  claim 4 , wherein the antagonist of an FGF receptor is SU5402 or PD173074. 
   
   
       14 . The method of  claim 3 , wherein the cells are provided as a blastocyst. 
   
   
       15 . A method of culturing pluripotent cells, comprising culturing the pluripotent cells in a culture medium comprising a MEK inhibitor and a GSK3 inhibitor, wherein the pluripotent cells undergo self-renewal. 
   
   
       16 . The method of  claim 15 , wherein the culture medium further comprises an antagonist of an FGF receptor. 
   
   
       17 . The method of  claim 15 , wherein the cells comprise mouse cells. 
   
   
       18 . The method of  claim 15 , wherein the cells comprise human cells. 
   
   
       19 . The method of  claim 15 , wherein the cells comprise rat cells. 
   
   
       20 . The method of  claim 15 , comprising culturing the cells in the absence of serum. 
   
   
       21 . The method of  claim 15 , wherein the MEK inhibitor is a MEK1 inhibitor. 
   
   
       22 . The method of  claim 15 , wherein the MEK inhibitor is selected from PD184352 and PD98059. 
   
   
       23 . The method of  claim 15 , wherein the GSK3 inhibitor is an inhibitor of GSK-3β. 
   
   
       24 . The method of  claim 23 , wherein the GSK3 inhibitor is CHIR99021. 
   
   
       25 . The method of  claim 16 , wherein the antagonist of an FGF receptor is SU5402 or PD173074.

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