改进的EBSS培养液与常用商品化试剂对鼠胚体外发育的影响

饶金鹏 金敏 金帆

[摘要] 目的 利用改进后的EBSS（SIGMA）培养液和常用商品化培养液G1/G2（Vitrolife），Quinns1026/Quinns1029（SAGE）对昆明系小白鼠胚胎进行体外培养，以对新建IVF实验室进行评估。 方法 对简单培养液EBSS进行改进，分别制得卵裂培养液和囊胚培养液以构成序贯培养系统。卵裂培养液： EBSS中加入适当浓度的丙酮酸钠，乳酸钠以及5种非必须氨基酸；囊胚培养液： EBSS中加入适当浓度的5种非必须氨基酸及6种必须氨基酸。将胚胎分成4组，A组使用EBSS （Earles Balanced Salt Solution）培养液，B组使用改进后的EBSS培养液，C组使用G1和G2培养液，D组使用Quinns1026和Quinns1029培养液，四种培养液均添加10%人血清白蛋白。结果 72 h后，鼠胚总体囊胚形成率为70.57%（614/870），其中A组的囊胚形成率为26.21%（27/103），B组为72.22%（143/198），C组为73.81%（155/210），D组为80.50%（289/359）B，C，D 3组的囊胚形成率显著高于A组（P<0.001）。 结论 通过鼠胚体外培养，对新建试管婴儿实验室进行了较好的质控检测，经改进后的EBSS培养液在鼠胚囊胚形成率上与商品化序贯培养液G1/G2，Quinns1026/Quinns1029相近，均远高于简单培养液EBSS，但前者成本较之常用商品化试剂有大幅度的降低，说明改进的EBSS培养液在鼠胚体外培养上具有较好的实用价值。

[关键词] EBSS；培养液；鼠胚培养；囊胚；质量控制

[中图分类号] R4 [文献标识码] A [文章编号] 1674-0742（2015）05（b）-0007-03

Effects of Improved EBSS Culture Media on the Development of Mouse Embryos in Vitro Compared with Common Commercial Media

RAO Jin-peng1，JIN Min1，JIN Fan2

1.Centre for Reproductive Medicine， The Second Affiliated Hospital， Zhejiang University， School of Medicine， Hangzhou， Zhejiang Province， 310052 China；2.Women's Hospital， School of Medicine， Zhejiang University， Hangzhou， Zhejiang Province， 310006 China

[Abstract] Objective Using improved EBSS（SIGMA） culture media and common commercial media G1/G2（Vitrolife） and Quinn's1026/Quinn's1029（SAGE） to culture mouse embryo of Kunming species in vitro for the assessment of a newly constructed IVF laboratory. Methods Modifying simple culture media EBSS by adding proper concentration of sodium pyruvate， sodium lactate and 5 nonessential amino acids to make cleavage stage media and adding proper concentration of 5 nonessential amino acids and 6 essential amino acids to make blastocyst stage media. 2-cell mouse embryos with normal morphology were collected and divided into 4 groups. Group A was cultured in EBSS （Earle's Balanced Salt Solution）， Group B was cultured in improved EBSS media， group C was cultured in G1/G2， group D was cultured in Quinn's1026/Quinn's1029， and each media was added with 10% human serum albumin （HSA）. Results The general blastocyst formation rate of mouse embryo at 72 h was 70.57%（614/870）. The blastocyst formation rate of group A was 26.21%（27/103）， that of group B was 72.22%（143/198）， that of group C was 73.81%（155/210） and that of group D was 80.50%（289/359）. Compared with group A， the blastocyst formation rate of group B， C and D was significantly higher， respectively （P<0.001）. Conclusion The mouse embryo culture in vitro served as a target to assess the quality control for a newly constructed IVF laboratory. Using improved EBSS media achieved similar mouse blastocyst formation rate compared with the common commercial media G1/G2 and Quinn's1026/Quinn's1029， which were significantly higher than simple media EBSS. However， the cost for improved EBSS media was much lower than common commercial media which means the improved EBSS media was superior for the development of mouse embryos in vitro.