一种改进的原代神经元的制备方法  

A Modified Protocol for the Preparation of Primary Neurons

作　　者：邓小云[1] 杨眉[1] 蔡芳[1] 潘乾[1] 龙志高[1] 邬玲仟[1] 梁德生[1] 戴和平[1] 夏昆[1] 夏家辉[1] 张灼华[1] 

机构地区：[1]中南大学医学遗传学国家重点实验室,中国湖南长沙410078

出　　处：《激光生物学报》2005年第2期145-149,共5页Acta Laser Biology Sinica

基　　金：国家自然科学基金资助项目(30170454;30123006;30300101)

摘　　要：神经元是神经组织的结构和功能单位。从胚胎中分离出神经元时,由于它们在原位组织中完成了分裂和分化,原代培养的神经元将不会分裂、增殖。18d孕龄的SD大鼠胎鼠脑组织中分离原代神经元的过程中,采用了改进的细胞分离方法。分离的原代细胞经免疫荧光实验证实含有大量的神经元。pEGFP质粒转染和细胞免疫荧光实验结果显示,转染的原代神经元中GFP报告基因有较高的表达,这表明,原代培养的神经元适宜于后续实验的进行。在对改进型方法和传统型方法的细胞分离效果比较时,发现改进型方法在消化过程中产生的gDNA絮状物较传统型的少,接种后发现细胞分散均匀,无杂物。所获得的原代细胞的总数比传统型方法多出大约23%。Neuronal cell is the structural and functional unit of the nerve tissues. Neurons were highly differentiated when they were dissected from the embryonic brain. As a result they cannot mitose and proliferate after dissected and seeded in the plates. Primary neurons were prepared from embryonic day 18 rat embryos. The cellular immunofluorescence showed that there exist a lot of neuronal cell in the cultures. The reporter gene of GFP can strongly express in the neuronal cell identified by the plasmid transferction and cellular immunofluorescence. The modified protocol for the preparation of neurons can improve the total number of the cell to about 23 percent.

关 键 词：原代细胞培养 神经元细胞 细胞免疫荧光 

分 类 号：Q813.1[生物学—生物工程]