昆明小白鼠胚胎干细胞分离培养的影响因素

Factors affecting the isolation and culture of embryonic stem cell in KM mice

为了研究影响胚胎干细胞（ESC）体外分离克隆的因素,试验采用3~5日龄大鼠心肌细胞制成心肌细胞条件培养基（RH-CM）,比较了4种培养体系对小鼠胚胎干细胞体外培养的影响,并分析比较了不同饲养层对胚胎干细胞分离克隆造成的影响。结果表明：添加重组鼠白血病抑制因子和心肌细胞条件培养基在胚胎贴壁率、内细胞团（ICM）形成率和原代胚胎干细胞集落形成率以及抑制分化的过程中起到的作用显著优于未添加任何诱导成分的培养基;作为饲养层,小鼠胚胎成纤维细胞（MEF）分离培养胚胎干细胞的效果优于小鼠睾丸支持细胞（MS）。

The experiment was designed to study the influencing factors of isolation and culture of KM mouse embryonic stem cell （ESC）. In this experiment, conditioned medium was made by cadioeyte from 3- 5 d rat for the first time, effect of four culture system and different feeder layers on isolation and culture of ESC were studied. The result showed that culture medium adding LIF or RH - CM had a significant difference in adherent ratio, ICM formation ratio, primary ESC formation and inhibiting differentiation comparing with the culture medium without LIF or RH -CM, feeder layer made by MEF had a better effect of isolation and culture of ESC than feeder layer made by mouse sertoli cells.