慢病毒转染和电转染原代C57BL/6小鼠神经干细胞的比较

Comparison of lentivirus transfection and electroporation transfection in primary neural stem cells of C57BL/6 mice

摘要:

摘要：目的  比较慢病毒与电转染两种技术转染小鼠原代神经干细胞的优劣，获得神经干细胞高效转染的方法。方法  从C57BL/6小鼠脑组织中提取神经干细胞，进行原代培养；分别进行慢病毒转染和电转染，通过观察绿色荧光蛋白在细胞中的表达，比较两种转染方法的转染效果，并用噻唑蓝（MTT）法计算相对存活率，比较两种方法转染后对细胞的影响。结果  慢病毒与电转染两种转染技术转染神经干细胞48 h后，慢病毒的转染率为（77.6±6.6）%，高于电转染法转染率[（29.2±4.8）%]，两者比较，差异有统计学意义（P <0.05）；转染72 h后，慢病毒转染的细胞荧光表达量为（60.0±3.6）%，电转染细胞的荧光表达量仅剩（12.8±2.4）%；MTT法结果显示，电转染组细胞相对存活率为（75.8±2.3）%，慢病毒转染组细胞相对存活率为（70.1±1.4）%，两者比较，差异无统计学意义（P >0.05）。结论  对于原代神经干细胞转染，慢病毒转染法比电转染法具有更高的转染效率及长久稳定性，转染后细胞存活率与电转染法接近，因此慢病毒转染法更能满足研究者的需求。

Abstract:

Abstract: Objective To develop an efficient method to transfect mouse primary neural stem cells (NSCs). Methods Primary neural stem cells were isolated from brain tissue of C57BL/6 mice, then, lentivirus transfection and electroporation methods were used to transfect primary NSCs. By observing the green fluorescent protein expression the transfection effect of the two methods was compared. The survival rate of primary NSCs was assessed using MTT method, the effect of the two methods on primary NSCs were compared. Results The rate of lentivirus transfection [(77.6 ± 6.6)%] was higher than that of electroporation [(29.2 ± 4.8) %, P < 0.05] 48 h after transfection. After 72 h, the amount of green fluorescence expression was (60.0 ± 3.6) % by lentivirus transfection and only (12.8 ± 2.4) % by electroporation method. The survival rate of lentivirus transfection [(70.1 ± 1.4) %] was not significantly lower than that of electroporation [(75.8 ± 2.3) %, P > 0.05]. Conclusions Lentivirus transfection is more efficient than electroporation for NSCs.