小鼠EG细胞体外分化为神经细胞的一种简单有效方法  

A simple and effective method differentiating EG cells into nerve cells in vitro in mice

作　　者：郭志林[1] 王新庄 张涌[1] 王木[2] 王轶敏 胡文举[1] 

机构地区：[1]西北农林科技大学生物工程研究所,陕西杨凌712100 [2]河南省肉牛工程技术研究中心,河南郑州450003

出　　处：《西北农林科技大学学报（自然科学版）》2008年第5期48-52,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：河南省杰出人才创新基金项目(0421002100)

摘　　要：【目的】探讨一种简单有效的小鼠EG（Embryonic germ cells,EG）细胞体外分化为神经细胞的方法。【方法】将昆明小鼠EG细胞在老化饲养层上培养3-4周,使EG细胞在高密度条件下向神经细胞分化,研究不同浓度维甲酸（Retinoic acid,RA）对EG细胞分化的诱导效果。【结果】小鼠EG细胞在高密度培养条件下向神经细胞分化,可获得较高比例的神经细胞样克隆,nestin染色阳性率达62.9%;经检测EG细胞分化产生的神经细胞样克隆外层细胞,以及从克隆中迁出的细长细胞均表达神经前体细胞标志nestin;在相同RA浓度下,随着EG细胞分化时间的延长,TH^＋（tyrosine hydroxylase,TH）神经细胞的比例增加;而在不同RA浓度下,随着RA浓度的升高,TH^＋神经细胞的比例呈增加趋势,经RA诱导14 d,最高可获得（2.3±0.2）%的TH^＋神经细胞。【结论】昆明小鼠EG细胞在老化饲养层上高密度培养条件下,能自发分化为神经细胞,RA诱导可显著提高分化细胞群体中TH^＋神经元细胞的比例。试验建立的这种简单有效方法可用于EG细胞体外分化为神经细胞。[Objective] The objective of the present study was to establish a simple and effective method differentiating EG （Embryonic germ cells,EG） cells into nerve cells in vitro in mice. [Method] Neuron differentiation of mice EG cells was started when EG cells derived from Kunmingbai Str. Mice were cultured on aging feeder to form high-density culture after 3-4 weeks,which was a method of induction and differentiation of neuron not by the phase of embryoid bodies forming. [Result] The results showed that this method got higher proportional neuron-like clone （62. 9% （78/124））after differentiation for some time,Enveloping cells of neuron-like clone derived from EG cells and slender cells emigrating from clone both expressed nestin marking of neuronal precursor cells;when straggling neural cells clone was induced by different concentration RA （Retinoic acid,RA） ,the proportion of TH＋ （tyrosine hydroxylase,TH）cells increased with the increase of differentiation time of EG cells in a same concentration RA; The proportion of TH^＋ cells took on a tendency of increase with the rise of concentration RA in different concentration RA;The highest percentage of （2.3±0.2）% TH^＋ neural cells was achieved by the induction for 14 d. [Conclusion] Above-mentioned results showed that EG cells derived from Kunming Str. Mice could spontaneously differentiate into neural cells cultured with high concentration on ageing feeder layer,which was a simple and effective method differentiating mice EG cells into nerve cells.

关 键 词：EG细胞 神经细胞 体外分化 昆明小鼠 

分 类 号：S814.8[农业科学—畜牧学] Q954.4[农业科学—畜牧兽医]