嗅球神经元的体外培养研究

Study on Cell Culture of Olfactory Bulb Neurons in Vitro

目的 :建立稳定的、产出率高的嗅球神经元离散细胞培养系统。方法 :取出生后5～6d大鼠的嗅球于体外培养。48h后随机分为对照组和阿糖胞苷(Ara -C)组连续培养14d。动态观察细胞生长情况 ,进行免疫细胞化学染色。结果 :嗅球神经元可以在体外培养成功 ,且对神经元有特异性的标志蛋白抗神经丝蛋白 (NFP)呈阳性反应。加入抗有丝分裂剂Ara-C后2组标本计数第1d差别无统计学意义 (P>0.05) ;第5,7,14dAra-C组较对照组明显增高(P<0.01)。其神经突生长指数随着时间的延长而增长 ,于7d后逐渐下降。结论 :在有血清的培养条件下 ,离散后的嗅球神经元仍可在体外存活 ,并有神经突生长、标记蛋白的表达。为生理学、分子生物学及药理学研究提供了一个有用的实验模型。

Objective:To establish a steady cell culture model of olfactory bulb neurons in vitro.Methods:The olfactory bulbs of5～6-day postnatal Wistar rats were cultured in vitro.After48h,the samples were randomly divided into control group and cytosine arabinoside(Ara-C)group.The neurons were cultured for14days.Results:The olfactory bulb neurons could survive in vitro.The immunoreactivity of neurofilament proˉtein(NFP)(a specific marker protein for neurons)was detected in the cultured neurons.After the Ara-C(an antimitotic drug)was added to medium,no difference of neuron count between the two groups could be found1-day later(P>0.05).However,after5,7and14days culture,the neuron counts in the Ara-C group draˉmatically increased as compared with those in the control group(P<0.01).The neuritogenesis index increased as time went on,but it gradually decreased after7days culture.Conclusion:The dissociated olfactory bulb neurons cultivated in the medium with fetal bovine serum survived with normal cell phenotype,neurite growth and the expression of neurifilament protein.The neuronal culture is a useful model for physiological,molecuˉlo-biological and pharmacological studies.