小鼠附植前胚胎培养基的优化

从4种常用培养基M16、MTF、CZB、KSOM中筛选适合CD-1小鼠附植前胚胎发育的体外培养基,并分析磷酸二氢钾、乳酸钠、葡萄糖、牛磺酸以及必需与非必需氨基酸对CD-1小鼠胚胎体外培养的影响。发现KSOM的培养效果明显优于其他培养基。在KSOM中添加必需氨基酸对8-细胞以前的胚胎发育有抑制作用,但添加牛磺酸、必需氨基酸与非必需氨基酸都可提高囊胚的细胞数。优化后的分阶段KSOM培养基显著提高了早期胚胎的囊胚发育率和囊胚期细胞数目。

Optimization of Mouse Preimplantation Embryo Culture Media

To develop an effective CD-1 mouse embryo culture medium,we evaluated the effects of several culture media including M16,MTF,CZB,KSOM and taurine,essential amino acids and non- essential amino acids were added in KSOM medium to observe their effects on the development of mouse preimplantation embryos.The KSOM was anout standing one among the media listed.Adding taurine,essential amino acids and nonessential amino acids in KSOM medium can increase the cell number of CD-1 mouse morula and blast ocysts,however,adding essential amino acids in KSOM media retarded the devel opmentrate of early CD-1 mouse embryos.Staged KSOM media K1/K2(K1 for pre-8-cell stage embryos containing 0.175 mM KH₂PO₄,20 mM sodium lactate,0.1mM glucose and 0.1mM taurine,K2 for post-8-cell stage embryos containing 0.35 mM KH₂PO₄,5mM sodiumlactate,5.56 mM glucose,0.1 mM taurine,0.25 mL 100× nonessential amino acids and 0.5 mL 50×essential amino acids)were formed by changing the dose of sodium lactate,KH₂PO₄,glucose,taurine,essential amino acids and nonessential amino acids.The optimized K1/K2 media could significantly improve the blast ocysts developmentrate and the cell number of morula and blast ocysts.