DEVELOPMENT OF PRE-IMPLANTATION EMBRYOS OF THE GOLDEN-HAMSTER IN A DEFINED CULTURE-MEDIUM
DEVELOPMENT OF PRE-IMPLANTATION EMBRYOS OF THE GOLDEN-HAMSTER IN A DEFINED CULTURE-MEDIUM
复制标题
DOI：
10.1095/biolreprod28.1.235
复制
发表时间：
1983-01-01
影响因子：
3.6
通讯作者：
LIEBERMAN, G
中科院分区：
文献类型：
--
作者：
BAVISTER, BD;LEIBFRIED, ML;LIEBERMAN, G
用孕鼠血清促性腺激素（PMSG）和人绒毛膜促性腺激素（hCG）超数排卵的金黄地鼠获得8细胞胚胎。将胚胎在设计用于仓鼠卵母细胞体外受精的限定培养基（改良Tyrode溶液）中培养24或32 h。在某些实验中，该培养基补充了氨基酸（谷氨酰胺、苯丙氨酸、甲硫氨酸和异亮氨酸）和维生素（Spille最低必需培养基维生素补充剂）。在培养期结束时，记录发育至囊胚期的胚胎数量。在其他实验中，检查了改变培养基的渗透压（225、250、275和300 mOsmol/kg）和pH（6.8和7.4）对胚泡形成的影响。发现早期8-细胞胚胎的能力（约100%）与早期8-细胞胚胎的能力（约100%）之间存在显著差异。卵激活后54小时）和晚期8细胞胚胎（约10小时）。卵活化后62小时）在培养物中发育。在未补充的培养基中，只有2%的早期8-细胞胚胎发育到囊胚期，而晚期8-细胞胚胎发育到囊胚期的比例为22%。这4种氨基酸对胚胎发育有显著影响，在氨基酸存在下，36%的早期8细胞胚胎发育成囊胚（增加18倍）。氨基酸还使发育成囊胚的晚期8细胞胚胎的百分比从22%增加到66%。在仓鼠胚胎发育的8细胞阶段的关键时期，可能会发生重要的代谢变化。当培养基中含有维生素时，未观察到对发育的额外影响。当由培养的8-细胞胚胎形成的胚泡通过手术转移到假孕仓鼠时，25%发育成正常闭锁胎儿，5个正常外观的幼崽出生，其中4个存活。这些结果代表了一种实现仓鼠胚胎体外完整植入前发育的方法。
Eight-cell embryos were recovered from mated golden hamsters that had been superovulated with pregnant mare''s serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG). Embryos were cultured for 24 or 32 h in a defined medium (modified Tyrode''s solution) designed for fertilization of hamster oocytes in vitro. This medium was supplemented in some experiments with amino acids (glutamine, phenylalanine, methionine and isoleucine) and with vitamins (Eagle''s Minimum Essential Medium vitamin supplement). At the end of the culture period, the numbers of embryos developing to the blastocyst stage were recorded. In other experiments, the effects of varying the osmotic pressure (225, 250, 275 and 300 mOsmol/kg) and the pH (6.8 and 7.4) of the culture medium on blastocyst formation were examined. A differece was found between the ability of early 8-cell embryos (.apprx. 54 h post-egg activation) and late 8-cell embryos (.apprx. 62 h post-egg activation) to develop in culture. In the unsupplemented culture medium, only 2% of early 8-cell embryos developed to the blastocyst stage compared with 22% of late 8-cell embryos. A marked effect of the 4 amino acids on development was found. In the presence of amino acids, 36% of early 8-cell embryos developed into blastocysts (18-fold increase). The amino acids also increased the percentage of late 8-cell embryos that developed into blastocysts from 22 to 66%. An important metabolic change may occur in hamster embryos during a critical period at the 8-cell stage of development. No additional effect on development was observed when vitamins were included in the culture medium. No significant effect of either osmotic pressure or pH of the culture medium on development was found. When blastocysts formed from cultured 8-cell embryos were transferred surgically to pseudopregnant hamsters, .apprx. 25% developed into normal-locking fetuses and 5 normal-looking young were born, 4 of which have survived. These results represent an approach towards achieving complete preimplantation development of hamster embryos in vitro.