小鼠胚胎体外培养方法的改良

孙芳园 段新崇 张振南 张晓 邸科前 李相运*

doi:10.16098/j.issn.0529-1356.2015.01.023

解剖学报 ›› 2015, Vol. 46 ›› Issue (1) : 127-132. DOI: 10.16098/j.issn.0529-1356.2015.01.023

技术方法

小鼠胚胎体外培养方法的改良

孙芳园¹ 段新崇¹ 张振南¹ 张晓¹ 邸科前² 李相运^1*

作者信息 +

Improvement of the method for in vitro culture of mouse embryos

SUN Fang-yuan¹ DUAN Xin-chong¹ZHANG Zhen-nan¹ZHANG Xiao¹ DI Ke-qian² LI Xiang-yun ^1*

Author information +

文章历史 +

摘要

目的改进胚胎培养系统以进一步提高胚胎质量与发育潜能。方法采用内径0.21mm、外径0.28mm的塑料毛细管培养胚胎，将吸有胚胎的毛细管浸在盛有蒸馏水的烧杯内，再将烧杯放置在培养箱里的磁力搅拌器上，允许胚胎在微小的空间内发育并伴随着水的旋转而摆动，更好地模拟了胚胎在输卵管中的运动环境。结果分别对85枚、82枚2-细胞胚胎进行毛细管及微滴培养，其中毛细管培养胚胎的囊胚率以及胚胎细胞数（85.4%，57.0）显著高于微滴培养（36.5%，20.6），囊胚率差异显著(P＜0.05)。且接种在Matrigel基底膜上形成的内细胞团集落显著增大。结论小鼠胚胎体外培养方法的改良——毛细管培养，促进小鼠植入前胚胎的体外发育。

Abstract

Objective To improve the embryo culture system for the quality of embryos and developmental potential. Methods In this study a rotating plastic capillary with an inside diameter of 0.21mm and an outside diameter of 0.28 mm were used to mimic oviduct properties and culture embryo in vitro. The capillary was inserted into a pipette tip, and then one to ten mouse 2-cell embryos in KSOM medium were drawn into a capillary by pipette. The capillary was immersed in a beaker containing a certain amount of autoclaved distilled water on a magnetic stirrer and then the device was placed in an incubator. After 48 hours culture the embryos were withdrew and their developmental potential were detected. Results Eighty-five and eight-two two-cells embryo were cultured in capillaries and microdrops, respectively. The blastocyst rate and average cell number are significantly higher in capillaries (85.4% and 57.0) than in microdrops (36.5%and 20.6). Capillary-cultured embryos formed bigger outgrowths than that of microdrop-cultured ones when they were seeded on matrigel-treated dishes. Conclusion Capillary culture, a method for in vitro culture the mouse embryo, raises success rates following human assisted reproduction.

导出引用

孙芳园段新崇张振南张晓邸科前李相运*. 小鼠胚胎体外培养方法的改良[J]. 解剖学报. 2015, 46(1): 127-132 https://doi.org/10.16098/j.issn.0529-1356.2015.01.023

SUN Fang-yuan DUAN Xin-chong ZHANG Zhen-nan ZHANG Xiao DI Ke-qian LI Xiang-yun*. Improvement of the method for in vitro culture of mouse embryos[J]. Acta Anatomica Sinica. 2015, 46(1): 127-132 https://doi.org/10.16098/j.issn.0529-1356.2015.01.023

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