Pluripotent Cells From Rat and Other Species

Abstract

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

1 - 160 . (canceled) 
     
     
         161 . A genetically engineered rat obtained by a method comprising
 genetically modifying a rat pluripotent cell expressing two or more of Nanog, Oct4, FGF4, and Sox-2; and   introducing the pluripotent cell into a rat embryo to produce a genetically modified rat, wherein the rat pluripotent cell contributes to the germline of the genetically modified rat.   
     
     
         162 . The genetically engineered rat of  claim 161 , wherein the rat pluripotent cell expresses Nanog, Oct4, and Sox-2. 
     
     
         163 . The genetically engineered rat of  claim 161 , wherein the rat pluripotent cell further expresses alkaline phosphatase. 
     
     
         164 . The genetically engineered rat of  claim 161 , wherein the rat pluripotent cell expresses Rex1, Stella, FGF4, and Sox-2. 
     
     
         165 . The genetically engineered rat of  claim 161 , wherein the rat pluripotent cell does not express FGF5. 
     
     
         166 . The genetically engineered rat of  claim 161 , wherein the rat pluripotent cell is capable of forming a teratoma or teratocarcinoma in which differentiated cells from all three germ layers are present. 
     
     
         167 . The genetically engineered rat of  claim 161 , wherein the rat pluripotent cell is capable of growth and/or proliferation as a single cell in culture. 
     
     
         168 . The genetically engineered rat of  claim 161 , wherein the pluripotent rat cell is derived from a blastocyst by a method comprising:
 culturing the blastocyst in the presence of a MEK inhibitor and a GSK3 inhibitor, to obtain an inner cell mass;   isolating and dissociating the primary outgrowths of the inner cell mass;   isolating a cell or cells from the dissociated primary outgrowths of the inner cell mass; and   culturing the isolated cell or cells in the presence of a MEK inhibitor, a GSK3 inhibitor, and an antagonist of an FGF receptor.   
     
     
         169 . The genetically engineered rat of  claim 168 , wherein the pluripotent rat cell expresses one or more of Nanog, Oct4, FGF4, Sox-2, and alkaline phosphatase. 
     
     
         170 . A genetically engineered rat obtained as the progeny of the genetically engineered rat of  claim 161 . 
     
     
         171 . The genetically engineered rat of  claim 161 , wherein the genetically engineered rat is homozygous null for a gene of interest. 
     
     
         172 . The genetically engineered rat of  claim 161 , wherein the genetically modifying comprises replacing a gene in the rat with a corresponding human gene. 
     
     
         173 . The genetically engineered rat of  claim 161 , wherein the genetically modifying comprises:
 knocking out one or more genes of interest; or   targeted insertion of a gene of interest.   
     
     
         174 . The genetically engineered rat of  claim 161  for use in drug discovery and/or testing. 
     
     
         175 . The genetically engineered rat of  claim 161  for use as a model for human or animal disease. 
     
     
         176 . The genetically engineered rat of  claim 175  for use in assessments of behavioural recovery or cognitive repair.