| 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,+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+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+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. |
| 关键词: EG细胞 神经细胞 体外分化 昆明小鼠 |
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| 基金项目:河南省杰出人才创新基金项目（0421002100） |
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| A simple and effective method differentiating EG cells into nerve cells in vitro in mice |
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| Abstract: |
| 【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. |
| Key words: EG cell neural cell differentiation in vitro Kunming Str.Mice |
