昆明白小鼠1细胞胚胎体外培养系统的研究

DEVELOPMENT OF ALBINO KUNMING MOUSE 1-CELL EMBRYOS IN DIFFERENT CULTURAL SYSTEMS IN VITRO

研究发现在有或者没有磷酸盐的条件下，葡萄糖均抑制昆明白小鼠ｌ－４细胞期胚胎的体外发 育。在不含葡萄糖和磷酸盐的ＨＥＣＭ－ｌ中，桑椹率为４０．０５％（７４／１６８），而对照Ｇ－ＨＥＣＭ－１仅为 ８．１４％（７／８６）；不含葡萄糖含有磷酸盐的ＣＺＢ中，桑椹率为６７．１１％（９３／１５２），而对照ＴＡＬＰ仅 ６· ６７％（６／９０）。用不含葡萄糖而含有１． ０ｍｍｏ１／Ｌ谷氨酸肢和０． １１ｍｍｏｌ／Ｌ ＥＤＴＡ的ＣＺＢ液，与兔输 卵管上皮单层培养细胞（ＲＯＥＣ）协同培养小鼠１细胞胚，７３．３３％（１１０／１５０）胚胎发育至桑椹胚， 但没有观察到囊胚形成、用上述ＣＺＨＲＯＥＣ系统培养小鼠１细胞胚４８小时（３－４细胞），再移入 ＴＣＭ１９９＋１０％ＦＣＳ＋ＲＯＥＣ系统，有７６．７４％（６７／８６）胚胎发育至桑椹胚，９６／小时后，４０．７０％ （３５／８６）发育至囊胚。

The results of the experiments showed that in the medium containing phosphate or not, the presence of glucose for the first 48 hour culturing was detrimental to the development of 1-4 cell Kunming mouse embryos. The ratio of morula was 40. 05% (74/168) in the HECM-l lacking glucose and phosphate j 8. 14% (7/86) in the G-HECM-1 containing glucose and lacking phosphate; and 61. 18% (93/152) in the CZB medium, 6. 67% (6/90) in the control (TALP). The experimental material was cultured in the cultural system, in which the CZB medium containing lmmol/L glutamine and 0. 11mrnol/L EDTA but lacking glucose and co-cultured with ROEC (rabbit oviduct epithelium cells) allowed 73. 33% (110/ 150) of embryos to develop into morula stage, but blastocysts were not observed. As the material was co-cultured with ROEC using CZB medium for 48 hours, and then placed into ROEC +TCM199+ 10 %FCS for about 48 hours, 40. 70 % of 1-cell embryos developed into blastocysts.