CAJ | 학술논문

通过使用不同的卵母细胞采集方法、不同的培养液、培养时间和细胞因子对绵羊卵母细胞的体外成熟培养的影响进行了研究.结果表明：切卵法和抽吸法对卵母细胞成熟的影响前者优于后者,差异极显著(P<0.01)；培养液中添加FSH、LH 及FBS对绵羊卵母细胞的影响显著高于添加发情羊血清的且差异极显著(P<0.01)；绵羊的卵母细胞培养20 h的成熟率极显著低于培养22 h和24 h的成熟率(P<0.01),培养22 h和24 h的成熟率差异不显著(P>0.05),培养24 h和培养26 h的成熟率差异极显著(P<0.01),培养26 h卵母细胞老化对随后的体外受精不利；成熟液(培养液+FBS、LH、FSH、E2)中分别加入骨形态发生蛋白15(bone morphogenetie protein,BMP15)和成纤维细胞生长因子8 b(fibroblast growth factor 8b,FGF8b)2种细胞因子,成熟液中添加 BMP15组与添加FGF8b组卵母细胞的成熟率差异极显著(P<0.01),成熟液中添加 BMP15和对照组卵母细胞成熟率的差异极显著(P<0.01),加入 FGF8b组与对照组卵母细胞成熟率差异不显著(P>0.05).利用切卵法在培养液中加入 FSH、LHE2和细胞因子BMP15,培养22~24 h可以大大提高绵羊卵母细胞体外培养的成熟率.
통과사용불동적란모세포채집방법、불동적배양액、배양시간화세포인자대면양란모세포적체외성숙배양적영향진행료연구.결과표명：절란법화추흡법대란모세포성숙적영향전자우우후자,차이겁현저(P<0.01)；배양액중첨가FSH、LH 급FBS대면양란모세포적영향현저고우첨가발정양혈청적차차이겁현저(P<0.01)；면양적란모세포배양20 h적성숙솔겁현저저우배양22 h화24 h적성숙솔(P<0.01),배양22 h화24 h적성숙솔차이불현저(P>0.05),배양24 h화배양26 h적성숙솔차이겁현저(P<0.01),배양26 h란모세포노화대수후적체외수정불리；성숙액(배양액+FBS、LH、FSH、E2)중분별가입골형태발생단백15(bone morphogenetie protein,BMP15)화성섬유세포생장인자8 b(fibroblast growth factor 8b,FGF8b)2충세포인자,성숙액중첨가 BMP15조여첨가FGF8b조란모세포적성숙솔차이겁현저(P<0.01),성숙액중첨가 BMP15화대조조란모세포성숙솔적차이겁현저(P<0.01),가입 FGF8b조여대조조란모세포성숙솔차이불현저(P>0.05).이용절란법재배양액중가입 FSH、LHE2화세포인자BMP15,배양22~24 h가이대대제고면양란모세포체외배양적성숙솔.
The influences of the different methods for harvesting the occytes,different culture matura-tion medias and various culture time on IVM of sheep oocytes were studied in this paper.The results showed that the effects of slicing on oocyte maturation were superior to the aspiration,and there were most significant differences between them (P<0.01).The effects of FSH,LH,FBS were better than those of ESS (P<0.01).It was higher in group which the oocytes were cultured for 22 and 24 hours compared with group that cultured for 20 hours (P<0.05),but there were no differences between 22 hours and 24 hours (P>0.05).There were most siginificant differences between 24 h and 26 h (P<0.01).It was harmful that the culture time of occyte was 26 h for the subsequential IVF.BMP15 (bone morphogenetie protein)and FGF8b(fibroblast growth factor 8b)were adder to the matural media.The mutural solution was regarded as the control group.The group of adding BMP15 had higher rate of IVM than the group of adding FGF8b (P<0.01),and the group of adding FGF8b and the control group had no difference.There were most sig-nificant differences between the group of adding BMP15 and the control group (P<0.01).Using the meth-od of slicing and adding FSH,LH,E2,BMP15 in the culture solution,the rate of occyte maturation of IVM of the sheep was very high after culturing for 22-24h.