[Effect on development in NT embryos after transplantation of nuclei derived from transfected goat fetal fibroblasts suffering different treatments into enucleate eggs].
In order to improve the development rate of preimplantation nuclear transfer embryos (NT embryos) after transplanting nuclei derived from transgenic goat fetal cells, the donor fetal fibroblasts starved for 5 days in DMEM containing 0.5% FCS were divided into three groups and treated with different methods respectively before using as donor cell. Group 1 was frozen at -80 degrees C or in liquid nitrogen for several days or months. Group 2 was at first treated as the same as group 1, then cultured for 2-5 days in DMEM containing 10% FCS and starved for another 5 days subsequently. Group 3 was cultured for 2-5 days in DMEM containing 10% FCS and starved for another 5 days subsequently. The rate of G0/G1 phase cells from group 2 was 95.68% and significantly different from group 1's 88.66%. The rate of survival cells from group 2 was 99.9% and significantly different from group 1's 80.00% (P < 0.05).The morula- blastocyst stage NT embryos development rate of group 2 was 66.09% and significantly different from group 1's 22.00% and group 3's 50.51% (P < 0.05). All NT embryos of above three groups were transferred into synchronous oestrus recipients and the pregnant status of recipients was checked by B-mode ultrasound diagnosis after 35 days. The recipient pregnancy rate of group 2 was 45.83%, much higher than that of group 1(20.00%) and group 3 (29.58%). The result of this experiment showed that donor cells treated with freezing and two times starvation could significantly improve the rate of G0/G1 phase cells, the rate of survival cells, the NT embryos development rate and the recipient pregnancy rate.
Chen
Jian Quan
JQ
Shanghai Transgenic Research Center, Shanghai 201203.
Zhang
Ai Min
AM
Chen
Juan
J
Xu
Xu Jun
XJ
Liu
Guo Hui
GH
Zhu
Min
M
Liu
Ming Gang
MG
Cheng
Guo Xiang
GX
D000818:Animals; D001755:Blastocyst / Q000502:physiology; D004624:Embryo Transfer / Q000379:methods*; D005260:Female; D005333:Fetus / Q000166:cytology*; D005347:Fibroblasts / Q000166:cytology* / Q000502:physiology*; D006041:Goats; D053652:Nuclear Transfer Techniques; D011247:Pregnancy