Abstract: Objective To investigate the effect of mouse embryonic endoderm cells and chick embryo extract(CEE) on the primary culture of rabbit embryonic stem cell, Methods A new method have been used to study the establishment of rabbit embryonic stem cell line. The condition systems were made up by feeder and medium. Mouse embryonic endoderm cells were inactivated by mitomycin c as feeder. Some different mediums were tested in this system. The inner cell masses separated from rabbit blastocyst were put into this condition system. After the inner cell mass attached to the feeder layer and formed outgrowth，the outgrowth with embryonic stem cell’s typical morphology were passaged. Results In the system consisted of mouse embryonic endoderm cells inactivated as feeder, 78% D-MEM/F-12+20% Knockout SR+2 mmol/L L-Glutamine+1 %MEM NAA+0.1 mmol/L p -mercapto ethanol+8 ng/ml hrbFGF+2%CEE as medium, the rabbit inner cell masses could grew into outgrowth with embryonic stem cell’s typical morphology. Conclusions Endoderm cells maybe were better than embryonic fibroblast for rabbit embryonic stem cell growth, and CEE maybe contained some growth factor which could inhibit rabbit embryonic stem cell differentiation.

Key words: Rabbit embryonic stem cell, Establishment, Culture condition, Endoderm.