Effects of Recipient Oocyte and Embryo Culture System on Production of Hanwoo (Korean Native Cattle) Somatic Cell Nuclear Transferred Embryos

서지정보

ㆍ발행기관 : 한국동물번식학회 ㆍ수록지정보 : Reproductive & developmental biology / 32권 / 3호
ㆍ저자명 : Dong-Hoon Kim, Se-woong Kim, Min-Jung Lee, Seong-Hoon Bae, Gi-Sun Im, Hyun-Joo Lim, Byoung-Chul Yang, Hwan-Hoo Seong

목차

ABSTRACT INTRODUCTION MATERIALS AND METHODS Preparation of Bovine Oocytes Preparation of Somatic Cells Enucleation and Nuclear Transfer Fusion and Activation In Vitro Culture of Embryos Terminal Deoxynucleotidyl Transferase-Mediated dUTP Nick-End Labeling (TUNEL) Assay Statistical Analysis RESULTS Development of Hanwoo SCNT Embryos according to Recipient Oocyte Source Development of Hanwoo SCNT Embryos according to Culture Systems DISCUSSION REFERENCES

영어 초록

This study was conducted to investigate an effective recipient oocyte and culture system for producing of Hanwoo (Korean native cattle) somatic cell nuclear transfer (SCNT) embryos. Hanwoo ear skin fibroblasts were used as donor cells. In vitro matured Hanwoo or Holstein oocytes were enucleated, and single donor cells were transferred into the perivitelline space of the enucleated oocytes. The couplets were subsequently fused and activated. The reconstructed embryos were cultured in a conventional or sequential culture system. In the former, embryos were cultured in CR2aa medium for eight days; in the latter, embryos were cultured in modified CR2aa-A (mCR2-A) for three days and then further cultured in modified CR2aa-B (mCR2-B) for five days. In the experiment with the recipient oocyte, the rate of embryo development to the blastocyst stage was significantly (p<0.05) higher in Hanwoo recipient oocytes than in Holstein ones (48.8% vs 38.9%). BIastocysts derived from Hanwoo recipient oocytes contained significantly (p<0.05) higher numbers of total cells than those derived from Holstein recipient oocytes (156.0+-68.2 vs 134.7+-54.8)). There was no difference in the mean proportion of apoptotic cells in blastocysts between the sources of recipient oocytes. In the experiment with the embryo culture system, the blastocyst rate was somewhat higher in sequential system than in conventional system (50.0% vs 43.5%), though there was no significant difference. The numbers of total (160.0+-69.0 vs 156.7+-68.4) and apoptotic cells (14.0+-10.4 vs 11.8+-6.4)) were not different between the culture systems. In conclusion, the present study demonstrated that Hanwoo recipient oocytes and the sequential culture system were more effective in supporting the production of Hanwoo SCNT embryos.

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