[Isolation and nuclear transfer of ES-like cells colonies derived from embryos being cloning of bovine somatic].
In this experiment, it was designed to carry out proliferous culture of bovine blastocysts(day 7) derived from embryos cloned through bovine somatic cell nuclear transfer, isolating and passaging of ES cells. The cells of blastocysts, which were planted on feeder layer, formed small colonies within 24 h. The nest-shape colonies occurred after culturing for 2-3 days. After the colonies in the same shape were isolated and passaged 4-5 times, many different size colonies with monolayer of multi-cells appeared. The colonies that had been passaged 4-5 times were planted into 4-wells multi-dishes without feeder layer. The colonies with monolayer of multi-cells appeared after 24 h, spread all over the bottom of the dishes, emerged epidermis-like cells that appeared reticulate after 4-7 days. These cells were used as donor cells to carry out nuclear transfer. The results showed that 80% (40/50) of the reconstructed embryos cleaved, 5% (2/40) and 2.5% (1/40) of them developed to the morulaes and blastocyst stage, respectively. It revealed that ES-like cells derived embryos constructed through somatic cell nuclear transfer have the developmental potentials.
Dong
Ya-Juan
YJ
Center of Animal Embryo Engineering, Laiyang Agriculture College, Shandong, Laiyang 265200, China. ETcenter@lyac.edu.cn
Bai
Xue-Jin
XJ
Li
Jian-Dong
JD
Suzuki
Tatsuyuki
T
D000818:Animals; D001757:Blastomeres / Q000166:cytology; D002417:Cattle; D002455:Cell Division; D019976:Cloning, Organism; D004622:Embryo, Mammalian / Q000166:cytology*; D005314:Embryonic and Fetal Development; D053652:Nuclear Transfer Techniques