胚胎大鼠皮质神经元原代培养及神经元鉴定

The primary culture of embryonic rat cortical neurons and neurons identification

目的对原有的胚胎大鼠神经细胞原代培养进行改进,建立一种高效、稳定的的原代培养方法。方法自精确控制胎龄的大鼠胚胎大脑中分离出皮质,采用胰酶二次消化法制备神经细胞悬液。并选用神经元特异性烯醇化酶免疫荧光化学法对神经细胞进行鉴定。结果与单次消化法分离细胞及运用DMEM+10%胎牛血清重悬细胞进行种板的原方法相比,方法改进以后,细胞提取数量明显增加,且纯度较高。种板后细胞形态正常且能长期存活。结论该方法结果理想,简易高效,值得借鉴和推广。

Objective To establish a highly efficient and stable method for the primary culture by modified the origi- nal method of cultureing embryonic rat cortical neurons. Methods Cerebral cortex isolated from the precise control of gestational age was dissociated to single cells by the twice-enzyme isolation method. Immunofluoreseence of neuronspe cific enolase chemical methods was used to identify neurons. Results The impro-~ed method could markably increase the number of cells and the purity compared to the original method of single digestion method and suspending cells use the DMEM＋10~fetal bovine serum. The plateing neurons was eumorphism and has long-time livability. Conclusion The experimental method has a ideal result also is simple and efficient that is worth to reference and generalize.