小鼠1-细胞胚胎体外培养条件的研究

STUDIES ON THE CONDITIONS FOR THE CULTURE OF ONE CELL MOUSE EMBRYOS

目的 检验培养基（Ｗｈｉｔｔｅｎ、ＣＺＢ、ＳＯＭ 和ＫＳＯＭ）、培养器皿（进口、国产新旧平皿）与配制培养基的水质对小鼠胚胎体外发育的影响。 方法 采用微滴培养法培养昆明种小白鼠１－细胞胚胎，以胚胎发育到囊胚的比例为判断培养效果的标准。 结果 Ｗｈｉｔｔｅｎ、ＣＺＢ、ＳＯＭ 和ＫＳＯＭ 培养基中２－细胞发育率分别为１００％ 、１００％ 、１００％ 和９８．６％ ，而囊胚发育率却分别为０％ 、７８．３％ 、４．８％ 和９．５％ ；用三蒸水、五蒸水和去离子水配制的ＣＺＢ培养基，培养囊胚发育率分别为４０．８％ 、５４．１％ 和５２．４％ ；国产和进口新平皿的囊胚发育率分别为６８．０％ 和７２．２％ ；国产和进口旧平皿的培养囊胚发育率为２７．２％ 和４６．１％ 。 结论 昆明小白鼠早期胚胎存在体外发育阻断现象；ＣＺＢ培养基培养昆明小白鼠早期胚胎效果最好；用三蒸水、五蒸水和去离子水配制的ＣＺＢ培养基培养结果无显著差别；进口和国产新平皿的培养效果相近，但再次使用时培养效果显著下降。

Objective\ To study the effects of culture media, types of culture dish and types of water used for preparation of media on the in vitro development of one cell embryos in the mice of Kunming breed. Method\ One cell mouse embryos collected from the oviduct of the superovulated mice were cultured in microdrops of medium and the percentage of embryos developed to blastocyst stage was used as evaluation criteria. Results\ (1)When one cell mouse embryos were cultured in the Whitten's, CZB, SOM and KSOM media, the cleavage percentages were 100%,100%,100% and 98 6%, respectively, and the blastocyst percentages were 0%, 78 3%, 4 8% and 9 5%, respectively. (2)The blastocyst percentages obtained from the CZB media prepared from the triple distilled, fivefold distilled and deionized water were 40 8%, 54 1% and 52 4%, respectively. (3)The blastocyst percentages produced from the Chinese made and the imported culture dishes were 68 0% and 72 2%, respectively, but those obtained when the dishes were used for the second time were decreased significantly (being only 27 2% and 46 1%, respectively). Conclusions\ There existed a development block during the in vitro development of the mouse embryos of the Kunming breed. Among the media tested CZB was the best in supporting the development of one cell mouse embryos to blastocyst stage. No significant difference was found among the CZB media prepared from the triple distilled, fivefold distilled and deionized water in blastocyst percentages. Both the Chinese made and the imported culture dishes produced good results of embryo development when used for the first time but the blastocyst percentages decreased significantly when the culture dishes were washed and used for the second time.\;