运用无胰酶植块法培养大鼠嗅鞘细胞的研究

Study on the culture of olfactry ensheathing cells in rats with trypsin-free explant method

目的:探讨无胰酶植块法进行大鼠嗅鞘细胞(OECs)培养对OECs形态特征的影响。方法:选用妊娠14d左右的SD雌性大鼠,取出其胎鼠,剥离嗅球,剪碎组织,分别用常规培养法无胰酶植块法分离OECs,用含15%胎牛血清的DMEM/F12完全培养基培养嗅鞘细胞,免疫荧光染色检测p75神经生长因子受体(p75 NG-FR)、胶质纤维酸性蛋白(GFAP)的表达,同时测量0ECs轴突的长度。结果:无胰酶消化的OECs克隆团处有大量的OECs向外延伸,分离单个OECs发现与常规培养法相比,其神经细胞轴突显著伸长;常规培养法可获取1.04×10^7个细胞,无胰酶植块法可获取3.09×10^7个细胞。结论:无胰酶植块法可获取大量的OECs,且OECs的轴突较长。

Ohjective:To investigate the ffe nf trypsin-free explant methoul on the morphological charncteristice of rat olfactory ensheathing cells(OECs).Methods:Embryonic SD rats were separated from utero of female rats at about 14 days of pregnaney.The olfactory bulbs of embryonie SD rats were separated.OECs were isolated by conventional method and trypsin-free explant method respectively.The expression of p75 nerve growth factor receptor(p75 NGFR)and glial fibillary acidic protein(GFAP)was detected by immunofluoresence staining,The axon length of OECs was measured.Results:There are numerous OECs extending outward in the OECs clones with trypsin-free explant method.Compared with the conventional culture method,the axons of nerve cells with lrypsin-free explant method were signifi-canly extended.There were 3.09×10^7cells obtained with trypsinfre explant method,while 1.04×10^7cells were ob-tained with conventional culture method.Conclusion:Try psin-free explant method can obtain abundant OECs with lon-ger axons.