Effects of Different Donor Cells Treatment with Aphidicolin and Donor Cell Passages on the Preimplantation Development of Somatic Cell Nuclear Transfer Embryos in Minipig

doi:10.3969/j.issn.1674-5817.2011.05.007

Abstract

Abstract: Objective To optimize somatic cell nuclear transfer (SCNT) procedures in mini-pigs, the present study was designed to examine the effects of donor cell types, cell passage number and aphidicolin (APC) treatment on in vitro development of reconstructed embryos. Methods The Oviduct epithelial cells (OEC), ear fibroblast cells (EFC) and cumulus cells (CC) derived from mini-pigs were treated with serum starvation only or serum starvation followed by treatment of 0.1 ?g/ml APC. The reconstructed embryos were cultured for 7 days to evaluate their developmental competency. Results Cleavage and blastocyst formation rates of reconstructed embryos derived from the OEC by APC treatment were significantly higher than the serum starvation (61.82% vs 56.25%, 24.55% vs 17.86%; P<0.05). The cleavage rate from the EFC was significantly increased by APC treatment compared to serum starvation only (63.36% vs 57.01%; P<0.05). In the ooctyes with the CC, the reconstructed embryos could yield high blastocysts formation rate by APC treatment (29.63%; P<0.05). In the presence of APC, CC gave rise to highest cleavage and blastocyst formation rates among the three cell types. All of the three donors can be used as donor karyoplasts for nuclear transfer, and the efficiency is not influenced by culture passages. Conclusion The treatment of CC with serum starvation plus APC prior to nuclear transfer is more suitable in SCNT of mini-pigs.

Key words: aphidicolin, minipig, cell type, passage number, somatic cell nuclear transfer.

CLC Number:

Q95-33

ZHANG Ting-Yu, DAI Jian-Jun, ZHANG Shu-Shan, WU Cai-Feng, ZHANG De-Fu. Effects of Different Donor Cells Treatment with Aphidicolin and Donor Cell Passages on the Preimplantation Development of Somatic Cell Nuclear Transfer Embryos in Minipig[J]. Laboratory Animal and Comparative Medicine, 2011, 31(5): 338-344.

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