Han Myung-Sook, Niwa Koji
The Graduate School of Natural Science and Technology, Okayama University, Okayama, Japan.
J Reprod Dev. 2003 Jun;49(3):235-42. doi: 10.1262/jrd.49.235.
Rat 1-cell embryos, recovered from naturally mated females, were cultured in a chemically defined medium (mR1ECM) or in mR1ECM supplemented with BSA (4 mg/ml; mR1ECM-BSA) or fetal bovine serum (FBS; 10%, v:v; mR1ECM-FBS) instead of polyvinylalcohol. There was no difference in percentages of embryos that developed to the 2-cell to blastocyst stages between mR1ECM and mR1ECM-BSA, but in mR1ECM-FBS, no development beyond the 2-cell stage was observed. When embryos were transferred to mR1ECM-FBS from mR1ECM after 24 to 64 h of culture, development of embryos to and beyond the 4-cell stage was inhibited. However, when transferred after 80 h of culture, more embryos developed to blastocysts and hatching or hatched blastocysts than in embryos cultured in mR1ECM. When 8-cell embryos and early morulae obtained after 72 and 80 h of culture in mR1ECM, respectively, were cultured in mR1ECM-FBS, a higher proportion of early morulae developed to the blastocyst stage than did 8-cell embryos. When morulae obtained after culture in mR1ECM or mR1ECM-BSA were transferred to recipient females, there was no difference in proportions of fetuses obtained. However, a higher proportion of blastocysts cultured in mR1ECM-FBS developed to fetuses compared with those obtained in mR1ECM. These results indicate that BSA has neither deleterious nor beneficial effects on development of rat 1-cell embryos. In contrast, FBS has deleterious effects on early cleavage of embryos but it promotes more rapid development of morulae to blastocysts, resulting in better quality blastocysts.
从自然交配的雌性大鼠中回收的1细胞胚胎,在化学限定培养基(mR1ECM)中培养,或在添加牛血清白蛋白(4 mg/ml;mR1ECM-BSA)或胎牛血清(FBS;10%,v:v;mR1ECM-FBS)而非聚乙烯醇的mR1ECM中培养。mR1ECM和mR1ECM-BSA之间发育到2细胞至囊胚阶段的胚胎百分比没有差异,但在mR1ECM-FBS中,未观察到超过2细胞阶段的发育。当胚胎在培养24至64小时后从mR1ECM转移到mR1ECM-FBS中时,胚胎发育到4细胞及以上阶段受到抑制。然而,当在培养80小时后转移时,与在mR1ECM中培养的胚胎相比,更多的胚胎发育成囊胚和孵化或已孵化的囊胚。当分别在mR1ECM中培养72小时和80小时后获得的8细胞胚胎和早期桑椹胚在mR1ECM-FBS中培养时,早期桑椹胚发育到囊胚阶段的比例高于8细胞胚胎。当在mR1ECM或mR1ECM-BSA中培养后获得的桑椹胚转移到受体雌性大鼠中时,获得胎儿的比例没有差异。然而,与在mR1ECM中获得的囊胚相比,在mR1ECM-FBS中培养的囊胚发育成胎儿的比例更高。这些结果表明,牛血清白蛋白对大鼠1细胞胚胎的发育既无有害影响也无有益影响。相比之下,胎牛血清对胚胎的早期分裂有有害影响,但它促进桑椹胚更快地发育成囊胚,从而产生质量更好的囊胚。