成年小鼠海马神经细胞培养及其鉴定
Culture and Identification of Hippocampus Neurons in Adult Rats
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摘要: 目的 观察体外培养成年小鼠海马神经细胞生长情况及其形态学变化.方法 获取成年海马组织,制成细胞悬液,接种于培养板,分别于1,3,7,10,13,15 d观察细胞生长情况,用神经元特异烯醇化酶抗体经免疫组织化学染色技术鉴定神经元.结果 成年海马神经元接种后1~3 d,有较多的杂质和部分细胞漂浮,贴壁细胞较少量.每隔3 d半量换液后,杂质不断减少,但第1周细胞生长缓慢,第7~10 d才见细胞生长良好.培养15 d后,大部分细胞出现有明显的颗粒等早期退化征象.免疫组化染色证实培养细胞呈现神经元特异烯醇化酶染色阳性染色,免疫阳性产物主要分布于细胞质;证明是神经元.结论 体外培养成年海马神经元早期生长缓慢,7~10 d才显示良好的生长状态,2周左右细胞则开始退变.提示10 d以前的细胞是供体外研究用的理想细胞.Abstract: Objective To observe the morphology and characteristics of hippocampus neurons in adult rats in vitro.Method The hippocampus tissues from adult rats were collected,then digested into cell suspension by using 0.125% trypsin.Cell suspension was planted into wells and observed at day 1,3,7,10,13,and 15 after incubation.Neurons specific Enolase(NSE)antibody,recognized specifically NSE antigen in cultured cells,was used to identify neurons by SP-two-step staining method.Results During 1-3 days,the extensive bodi...
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Key words:
- Adult rats /
- Hippocampus neurons /
- NSE staining /
- Culture /
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