KSOM培养基营养成分调整对小鼠植入前期胚体外发育的影响

摘要/Abstract

摘要： 目的囊胚发育率低和发育迟缓是继“2-细胞阻滞”解决后小鼠胚胎体外培养中面临的两大问题。本实验通过对KSOM培养基营养成分的调整,观察对胚胎发育的影响,优化培养基的构成。方法采用微滴培养法培养昆明小鼠单细胞胚胎,以胚胎发育到囊胚的比例、孵出比例以及注射hCG后120h囊胚全细胞数作为判断培养效果的标准,比较KSOM在葡萄糖、牛血清白蛋白、氨基酸3种营养物质不同浓度下对小鼠植入前胚体外培养的效果。结果在未加氨基酸时葡萄糖、牛血清白蛋白的变化对注射hCG后120h囊胚形成率、囊胚全细胞数影响不大,添加氨基酸后,囊胚发育速度提高,部分孵出和全部孵出比例提高,囊胚全细胞数增大明显,且葡萄糖、牛血清白蛋白与氨基酸对囊胚的发育有共同促进的作用。结论通过添加适量的氨基酸、葡萄糖、BSA改良KSOM培养基能很好的促进小鼠植入前胚体外发育。

Abstract: Objective To solve the problem known as 2-cell block in in vitro culture of preimplantation embryos, modification of the nutritional components in potassium simplex optimized medium (KSOM) was attempted and the preimplantation development and cleavage of mouse embryos in such medium observed.Method One-cell mouse embryos collected from the oviduct of the superovulated mice were cultured in microdrops of different culture media for 5 days [144h after human chorionic gonadotrophin (hCG) injection], and the percentages of embryos developed to pre-blastocyst, unhatched blastocyst, partially hatched blastocyst and completely hatched blastocyst were recorded at deferent time points (96,120 and 144h post-hCG) and total cell number of blastocysts at 120 h post-hCG counted. With these indices as the evaluation criteria, we evaluated the modifications of the concentrations of glucose (0.2,5.56mmol/l), bovine serum albumin (BSA,1 and 4mg/ml), and amino acids (1/2X NEAA and EAA,or not) in KSOM for their effect on the development of Kunming mouse zygotes in vitro.Results Increasing the concentration of glucose and BSA was not shown to have significant effects on the rate of blastocyst formation at 120h post-hCG or the total cell counts in KSOM without amino acids. After supplementation of the medium with amino acids, the rate of partially and completely hatched blastocyts was significantly increased in KSOM or KSOM supplemented with high concentrations of glucose and BSA (KSOMGB). Glucose at the high concentration of 5.56mmol/L did not inhibit the development of mouse zygotes to hatched blastocysts cultured in KSOM supplemented with amino acids.Conclusion Increasing the concentration of glucose and BSA and amino acids in KSOM can promote the preimplantation development of mouse zygotes in in vitro culture.

中图分类号:

R-33
R321

黄吴键¹, 袁进¹, 邓星², 安靓¹. KSOM培养基营养成分调整对小鼠植入前期胚体外发育的影响[J]. 南方医科大学学报, 2005, 25(03): 256-261.

HUANG Wu-jian¹, YUAN Jin¹, DENG Xing², AN Jing¹. Effects of modification of the nutritional components in potassium simplex optimized medium on preimplantation development and cleavage of mouse embryos in vitro[J]. Journal of Southern Medical University, 2005, 25(03): 256-261.

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