摘要
目的 :探讨体外培养高纯度神经元的方法。方法 :取SD新生大鼠 ,胰蛋白酶消化、离心 ,原代培养出神经元和神经胶质的混合细胞 ;振摇法和胰蛋白酶消化法去掉小胶质和星形胶质 ,传代培养神经元 ,免疫细胞化学法鉴定培养神经元的纯度。结果 :第三次传代培养细胞 ,NF抗体标记阳性细胞遍布整张盖片 ,仅有零星散在CD68和GFAP阳性细胞。结论 :本实验传代、培养的细胞高纯度的神经元 。
Objective: To investigate the method of obtaining high content of neuron in vitro. Method: Cerebral tissue from newborn SD rats was digested with trypsin and centrifugalized. The mixture of neuron and neuroglia was primarily cultivated. Microglia and astrocyte were freed from mixed cultures by gently tapping and repeating trypsinization of the mixture, respectively. Cells were again seeded at least two times. Purity of cells was assessed by immunocytochemistry using anti-neurofilament(NF), CD68 antibodies( as microglial marker) and anti-glial fibrillary acidic protein(GFAP) antibodies. Results: NF positive cells spread and distributed densely throughout the coverslips. In contrast, there were sporadic and scattered CD68 and GFAP positive cells throughout the coverslips. Conclusion: The method that authors have adopted in the experiment can cultivate highly purified neuron, which underlies further investigation.
出处
《泸州医学院学报》
2003年第3期200-202,共3页
Journal of Luzhou Medical College
关键词
神经元
传代培养
纯化
Neuron
Cultivation
Purification
