小鼠早胚体外培养成分确定培养基的实验研究

Study on chemical defined medium for the culture of mouse early embryos

目的:建立小鼠早胚体外培养成分确定培养基,并与传统培养基对小鼠早胚发育的效果进行比较?方法:用添加不同浓度聚乙烯醇(polyvinyl alcoho,PVA)和不同浓度氨基酸的CZB培养基和添加输卵管细胞?胎牛血清(fetal cattle serum, FCS)或牛血清白蛋白(bovine serum albumin, BSA)培养基培养小鼠早胚,观察它们对囊胚发育的影响?结果:含0.05 mg/ml PVA?0.5×必需氨基酸(essential amino acids solution, EAA)和0.5×非必需氨基酸(non-essential amino acids solution, NEAA)的CZB培养基可有效防止胚胎粘连,其总囊胚和扩张囊胚发育率显著高于其余各氨基酸添加组和对照组?尽管该培养基的总体发育效果低于输卵管细胞添加组,但其总囊胚发育率与BSA添加组差异无统计学意义,而且明显高于胎牛血清添加组?其扩张囊胚发育率与胎牛血清添加组差异无统计学意义?结论:小鼠早胚在含0.05 mg/ml PVA和0.5×EAA及0.5×NEAA的成分确定的CZB培养基中具有与在BSA添加培养基中相同的囊胚发育能力,且具有与在血清添加培养基中相同的扩张囊胚发育能力?

Objective: To establish a chemical defined medium for the culture of mouse early embryos and to compare the effect of this medium with that of the traditional medium on in vitro development of mouse early embryos. Methods: Mouse early embryos were cultured in CZB media supplememted respectively with polyvinyl alcohol(PVA) at different concentrations, amino acid at different concentrations, oviduct cells, fetal cattle serum (FCS), and bovine serum albumin(BSA) to study the effects of different media on the development of blastocysts. Results: The medium supplemented with 0.05 mg/ml PVA, 0.5×EAA+0.5×NEAA can effectively prevent the early embryos from adhesion to each other. The rate of total blastocysts and expanded blastocysts in this medium was significantly higher than those of control and other groups with different supplement of EAA and NEAA. The rate of total blastocysts in this medium was significantly lower than that in the medium supplemented with oviduct cells, significantly higher than that in the medium supplemented with FCS and has no significant difference with that in the medium supplemented with BSA. Conclusion: The chemical defined medium supplemented with 0.05 mg/ml PVA and 0.5×EAA+0.5×NEAA can maintain the development of embryo in vitro to blastocyst and is useful for studying the developmental mechanism of early mouse embryo. [