[1]孔慧娟,孙莹璞#,耿嘉暄.小鼠胚胎干细胞克隆形成和传代的影响因素*[J].郑州大学学报(医学版),2012,(01):36.
 KONG Huijuan,SUN Yingpu,GENG Jiaxuan.Influencing factors in the establishment of mouse embryonic stem cells[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2012,(01):36.
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小鼠胚胎干细胞克隆形成和传代的影响因素*
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2012年01期
页码:
36
栏目:
论著
出版日期:
2012-01-20

文章信息/Info

Title:
Influencing factors in the establishment of mouse embryonic stem cells
作者:
孔慧娟孙莹璞#耿嘉暄
郑州大学第一附属医院生殖医学中心 郑州450052
Author(s):
KONG HuijuanSUN YingpuGENG Jiaxuan
Medical Reproductive Center,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052
关键词:
小鼠胚胎干细胞培养液内细胞团
Keywords:
mouseembryonic stem cellculture mediuminner cell mass
分类号:
R321
摘要:
目的:探讨昆明小鼠胚胎干细胞(ESC)分离后克隆形成和细胞传代的几种影响因素,对其特性作初步鉴定。方法:对比超排卵和自然受孕2种囊胚获取方法、囊胚培养液和高糖DMEM培养液2种培养液对其克隆形成和细胞传代的影响。对贴壁后形成的原代ESC克隆进行评分。稳定传代的一株ESC进行鉴定。结果:超排卵组、自然受孕组的内细胞团形成率(64.9% vs 68.2%)和原代ECS克隆形成率(10.1% vs 14.5%)比较,差异均无统计学意义(χ2=0.345和1.385,P=0.557和0.239)。囊胚培养液组24 h囊胚孵出率明显高于DMEM培养液组(56.8% vs 17.1%)(χ2=33.026,P=0.001),原代ESC克隆形成率明显低于DMEM培养液组(35.2% vs 15.9%)(χ2=9.021,P=0.002)。ICM的评分与ESC克隆传代能力相关(r=0.531,P<0.001),直径70~100 μm、周边规则、隆起明显的ICM,ESC克隆传代持久。结论:高糖DMEM培养液对克隆的分离和传代有利。对ICM贴壁后形成的ESC克隆进行评分有助于确定挑选克隆传代的时机。
Abstract:
Aim:To investigate several kinds of influencing factors of Kuming mice embryonic stem cells(ESC) isolation and to perform the preliminary identification.Methods:Two blastocystobtaining methods,and two kinds of mediums (HLM and DLM) were compared, respectively. And the inner cell mass(ICM) was scored. Moreover, the omnipotent identification was carried out in a ESC strain with stable passage.Results:The ICM formation rates of the superovulation group and natural conception group (64.9% vs 68.2%) as well as the colony formation rate of the primary generation ESC (10.1% vs 14.5%) were similar, with no significant differences(χ2=0.345 and 1.385,P=0.557 and 0.239). The 24hour hatching rate of the HLM group was significantly higher than the DLM group(56.8% vs 17.1%)(χ2=33.026,P=0.001). But the colony formation rate of the primary generation was significantly lower (35.2% vs 15.9%)(χ2=9.021,P=0.002).Scoring of ICM was related to the ability of ESC clones passaged(rs=0.531,P<0.001).The ICM with a diameter of 70~100 μm, regular boundary and obvious elevation could have more passages.Conclusion:The high glucose DMEM medium is beneficial to the isolation and passage of cell clones. Scoring of the ICM is helpful to determine the timing of selecting clones and passage.

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备注/Memo

备注/Memo:
*国家自然科学基金资助项目30771105 doi:10.3969/j.issn.16716825.2012.01.012 #通讯作者,女,1962年1月生,博士,主任医师,教授,研究方向:生殖医学,Email:syp2008@vip.sina.com
更新日期/Last Update: 1900-01-01