新生大鼠螺旋神经节细胞体外生长特点及脂质体介导的转染效率

The Growth Characteristic of Spiral Ganglion Cells from Newborn Rats and the Liposome-mediated EGFP Transfection Efficiency in Vitro

陈请国;褚汉启;王显红;陈金;李建玲;周良强;熊浩;王燕;李志勇;崔永华;

1:华中科技大学同济医学院附属同济医院耳鼻咽喉-头颈外科

2:咸宁学院附属第一医院耳鼻咽喉科

摘要
目的探讨螺旋神经节细胞(spiral ganglion cells,SGCs)体外生长规律及特点,并观察阳离子脂质体转染SGCs的情况。方法从出生34 d的SD大鼠耳蜗中分离螺旋神经节组织,消化后放在含10%胎牛血清的DMEM/F12中培养,第2 d换用含2%B27及5μmol/L阿糖胞苷的Neurobasal培养基纯化SGCs,取第4 d活性良好的细胞爬片后,通过免疫荧光法用NF-200抗体鉴定SGCs;另外用质粒pEGFP-C2联合Lipofectamine 2000转染SGCs,观察其转染效率及对细胞活性的影响。结果体外培养的SGCs胞体饱满透亮,折光性好,一般能存活2周左右,第34 d的SD大鼠耳蜗中分离螺旋神经节组织,消化后放在含10%胎牛血清的DMEM/F12中培养,第2 d换用含2%B27及5μmol/L阿糖胞苷的Neurobasal培养基纯化SGCs,取第4 d活性良好的细胞爬片后,通过免疫荧光法用NF-200抗体鉴定SGCs;另外用质粒pEGFP-C2联合Lipofectamine 2000转染SGCs,观察其转染效率及对细胞活性的影响。结果体外培养的SGCs胞体饱满透亮,折光性好,一般能存活2周左右,第37 d活性最好。细胞对NF-200染色阳性;约10%的SGCs能被Lipofectamine 2000转染,但小部分细胞轴突缩短,甚至漂浮起来。结论含B27的Neurobasal培养基能培养出活性良好的SGCs;脂质体和质粒pEGFP-C2能成功地转染SGCs,但转染效率较低,并在一定程度上影响细胞的活性。
关键词
螺旋神经节细胞;Lipofectamine 2000;转染;B27
基金项目(Foundation):
国家自然科学基金资助项目(No.30672307)
作者
陈请国;褚汉启;王显红;陈金;李建玲;周良强;熊浩;王燕;李志勇;崔永华;
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