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KMID : 1005220070220040245
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Journal of Embryo Transfer
2007 Volume.22 No. 4 p.245 ~ p.249
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Effect of Concentration and Exposure Duration of FBS on Parthenogenetic Development of Porcine Follicular Oocytes
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Kim Hyun-Jong
Kim Il-Hwa
Lim Kyung-Sun
Son Dong-Soo
Choe Chang-Yong
Choi Sun-Ho
Cho Sang-Rae
Lee Woon-Kyu
Kim Sung-Jae
Sang Byung-Don
Han Man-Hye
Ryu Il-Sun
Kim In-Cheul
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Abstract
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The aim of present experiment was to examine hatching rate as in vitro indicator of viability of porcine embryos before early stage embryo transfer such as zygotes or 2-cell stage embryos. Cumulus-oocyte complexes (COCs) collected from ovaries were matured in North Carolina State University 23 (NCSU-23) containing 10% porcine follicular fluid (pFF), 10 ng/ml epidermal growth factor (EGF), follicle stimulating hormone (FSH), luteinizing hormone (LH), and 1mg/ml cysteine. After 24 hours, the COCs were transferred to the same medium without hormones. After 65h of maturation, oocytes were exposed to phosphate buffered saline (PBS) with 7% ethanol (v/v) for 7 minutes, and then the oocytes were washed and cultured in tissue culture medium (TCM) 199 containing 5 ug/ml cytochalasin B for 5h at in an atmosphere of 5% and 95% air with high humidity. After cytochalasin B treatment, the presumptive parthenotes were cultured in porcine zygote medium (PZM)-5 and cleavage of the parthenotes was assessed at 72h of activation, Normally cleaved parthenotes were cultured for an additional 8 days to evaluate their ability to develop to blastocyst and hatching stages. The fetal bovine serum (FBS) were added at Day 4 or 5 with concentrations of 2.5, 5 or 10%. The blastocyst rates were ranged within in each treatment. However hatching rate was dramatically decreased in non-addition group. In this experiment, embryo viability in female reproductive tract may be estimated before embryo transfer with in vitro culture adding FBS by hatching ability.
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KEYWORD
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porcine, parthenotes, fetal bovine serum, in vitro culture, hatching rate
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