改進(jìn)的EBSS培養(yǎng)液與常用商品化試劑對鼠胚體外發(fā)育的影響

饒金鵬 金敏 金帆

[摘要] 目的 利用改進(jìn)后的EBSS（SIGMA）培養(yǎng)液和常用商品化培養(yǎng)液G1/G2（Vitrolife），Quinns1026/Quinns1029（SAGE）對昆明系小白鼠胚胎進(jìn)行體外培養(yǎng)，以對新建IVF實(shí)驗(yàn)室進(jìn)行評估。 方法 對簡單培養(yǎng)液EBSS進(jìn)行改進(jìn)，分別制得卵裂培養(yǎng)液和囊胚培養(yǎng)液以構(gòu)成序貫培養(yǎng)系統(tǒng)。卵裂培養(yǎng)液： EBSS中加入適當(dāng)濃度的丙酮酸鈉，乳酸鈉以及5種非必須氨基酸；囊胚培養(yǎng)液： EBSS中加入適當(dāng)濃度的5種非必須氨基酸及6種必須氨基酸。將胚胎分成4組，A組使用EBSS （Earles Balanced Salt Solution）培養(yǎng)液，B組使用改進(jìn)后的EBSS培養(yǎng)液，C組使用G1和G2培養(yǎng)液，D組使用Quinns1026和Quinns1029培養(yǎng)液，四種培養(yǎng)液均添加10%人血清白蛋白。結(jié)果 72 h后，鼠胚總體囊胚形成率為70.57%（614/870），其中A組的囊胚形成率為26.21%（27/103），B組為72.22%（143/198），C組為73.81%（155/210），D組為80.50%（289/359）B，C，D 3組的囊胚形成率顯著高于A組（P<0.001）。 結(jié)論 通過鼠胚體外培養(yǎng)，對新建試管嬰兒實(shí)驗(yàn)室進(jìn)行了較好的質(zhì)控檢測，經(jīng)改進(jìn)后的EBSS培養(yǎng)液在鼠胚囊胚形成率上與商品化序貫培養(yǎng)液G1/G2，Quinns1026/Quinns1029相近，均遠(yuǎn)高于簡單培養(yǎng)液EBSS，但前者成本較之常用商品化試劑有大幅度的降低，說明改進(jìn)的EBSS培養(yǎng)液在鼠胚體外培養(yǎng)上具有較好的實(shí)用價(jià)值。

[關(guān)鍵詞] EBSS；培養(yǎng)液；鼠胚培養(yǎng)；囊胚；質(zhì)量控制

[中圖分類號] R4 [文獻(xiàn)標(biāo)識碼] A [文章編號] 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.