干细胞条件培养液对小鼠体外成熟卵母细胞发育潜能的影响

Effect of conditioned medium of mesenchymal stem cells on developmental potential of mouse in vitro maturation oocytes

目的：探讨骨髓间充质干细胞（MSCs）条件培养液对小鼠体外成熟卵母细胞发育潜能的影响。方法：分离、培养小鼠MSCs,获得MSCs条件培养液;收集4~6周小鼠生发泡期（GV）卵母细胞,在MSCs条件培养液中体外成熟（IVM）;同时部分小鼠促排处理,收集体内成熟卵母细胞;体外受精（IVF）后,比较受精率、4-细胞形成率及囊胚形成率差异;Hoechst33342染色后观察内细胞团（ICM）和滋养层细胞形成情况,计数内细胞团数。结果：IVM卵母细胞IVF后的受精率、4-细胞形成率和囊胚形成率及囊胚内细胞总数与体内成熟组比较,无显著差异（P〉0.05）;IVM组各时期的胚胎形态正常,囊胚内细胞染色后的荧光图像与体内成熟组基本一致。结论：MSCs条件培养液能保持小鼠IVM卵母细胞良好的质量和发育潜能,是一种较理想的IVM培养体系。

Objective： To investigate the effects of conditioned medium（ CM） of mesenchymal stem cells（ MSCs） on developmental potential of the mouse IVM oocytes. Methods：Mouse MSCs were isolated and cultured to harvest CM. Immature germinal vesicle（ GV） stage oocytes were collected and cultured in the CM of MSCs. The embryonic development of IVM oocytes after IVF were surveyed. Meanwhile,some mice were stimulated ovulation. The fertility rate,4-cell rate and blastulation rate of IVM oocytes after IVF were compared. Blastocysts stained with hoechst33342 to evaluate the quality by observation formation of inner cell mass（ ICM） and trophoblast cell,and counting the number of ICM of blastocysts with a fluorescence microscope. Results： The fertility rate,4-cell formation rate,blastulation rate and the number of total cells of IVM group were not statistically different compared with the control group（ P〉0. 05）. The embryonic morphology of each period of IVM oocytes after IVF were normal. The fluorescence pictures of ICM of IVM group were consistent with the in vivo group. Conclusion：Conditioned medium of MSCs is an efficient IVM system,which could keep the embryonic potential developmental of mouse IVM oocytes.