兔胚胎干细胞的早期培养条件初步研究

›› 2009, Vol. 29 ›› Issue (1): 17-21.

兔胚胎干细胞的早期培养条件初步研究

1.新疆农业大学动物医学学院，乌鲁木齐，830052；2.上海交通大学医学院实验动物科学部，上海，200025

收稿日期:2008-05-12 出版日期:2009-01-31 发布日期:2009-01-31

Study on Original Culture Condition of Rabbit Embryonic Stem Cell

1. School of Animal Medicine, Xinjiang Agricultural University, Urumqi 830052, China; 2. Department of Laboratory Animal Sciences, Shanghai Jiaotong University School of Medicine, Shanghai 200025，China

Received:2008-05-12 Online:2009-01-31 Published:2009-01-31

摘要/Abstract

摘要： 目的探索小鼠胚胎内胚层细胞和鸡胚提取物（CEE）对兔胚胎干细胞早期培养的影响。方法用全新的方法对兔胚胎干细胞的建系条件进行探讨。将分离到的囊胚期内细胞团接种到丝裂霉素C灭活的小鼠胚胎内胚层细胞饲养层和几种不同的培养液构成的培养体系中，观察内细胞团的贴壁和生长状况，并对由内细胞团生长出的具有胚胎干细胞形态的细胞集落进行传代。结果以灭活的小鼠胚胎内胚层细胞作为饲养层，以78%D-MEM/F-12+20%Knockout SR+2 mmol/LL-Glutamine+1%MEMNAA+0.1mmol/L β-mercaptoethanol+8ng/ml hrbFGF+2%CEE做培养液，兔内细胞团可以很好的贴壁，生长成具有标准的胚胎干细胞（ESC）形态的细胞集落。结论小鼠内胚层细胞和CEE联合使用可以很好的支持原代兔胚胎干细胞（RESC）的生长，小鼠内胚层细胞和CEE中可能含有某些抑制RESC分化的因子。

关键词: 兔胚胎干细胞, 建系, 培养条件, 内胚层

Abstract: Objective To investigate the effect of mouse embryonic endoderm cells and chick embryo extract(CEE) on the primary culture of rabbit embryonic stem cell, Methods A new method have been used to study the establishment of rabbit embryonic stem cell line. The condition systems were made up by feeder and medium. Mouse embryonic endoderm cells were inactivated by mitomycin c as feeder. Some different mediums were tested in this system. The inner cell masses separated from rabbit blastocyst were put into this condition system. After the inner cell mass attached to the feeder layer and formed outgrowth，the outgrowth with embryonic stem cell’s typical morphology were passaged. Results In the system consisted of mouse embryonic endoderm cells inactivated as feeder, 78% D-MEM/F-12+20% Knockout SR+2 mmol/L L-Glutamine+1 %MEM NAA+0.1 mmol/L p -mercapto ethanol+8 ng/ml hrbFGF+2%CEE as medium, the rabbit inner cell masses could grew into outgrowth with embryonic stem cell’s typical morphology. Conclusions Endoderm cells maybe were better than embryonic fibroblast for rabbit embryonic stem cell growth, and CEE maybe contained some growth factor which could inhibit rabbit embryonic stem cell differentiation.

Key words: Rabbit embryonic stem cell, Establishment, Culture condition, Endoderm.

张慧1，方贞付2，况玲1，陈学进2. 兔胚胎干细胞的早期培养条件初步研究[J]. , 2009, 29(1): 17-21.

ZHANG Hui1，FANG Zhen-fu2，KUANG Ling1，CHEN Xue-jin2. Study on Original Culture Condition of Rabbit Embryonic Stem Cell[J]. , 2009, 29(1): 17-21.