基质细胞衍生因子-1α-层粘连蛋白相互作用在体外诱导神经干细胞迁移和分化
[Stromal cell derived factor-1α-laminin crosstalk induced neural stem cells migration and differentiation in vitro].
作者信息
Zhang Jianqi, Li Xiaohong, Zhang Sai, Mei Jinping, Yang Qin, Zhang Juan
机构信息
Department of Cardiovascular Medicine, Affiliated Hospital of Logistics College of Armed Police Force, Tianjin 300162, China. Corresponding author: Zhang Jianqi, Email:
出版信息
Zhonghua Wei Zhong Bing Ji Jiu Yi Xue. 2017 Jan;29(1):57-60. doi: 10.3760/cma.j.issn.2095-4352.2017.01.012.
OBJECTIVE
To investigate the role of stromal cell derived factor-1α (SDF-1α)-laminin (LN) crosstalk in migration and differentiation of neural stem cells (NSCs).
METHODS
Original generation of NSCs collected from 14-day pregnant fetal rats were separated and cultivated, and the phenotype characteristics were identified with immunofluorescence. The third generation of NSCs were collected, and they were divided into four groups: poly-L-lysine (PLL) group, PLL+SDF-1α group, LN group and LN+SDF-1α group. The NSCs in PLL and LN groups were inoculated into plates coated with 0.1 mg/mL PLL or 1 mg/mL LN, and the NSCs in PLL+SDF-1α and LN+SDF-1αgroups were inoculated into plates coated with PLL or LN and 1 mg/mL SDF-1α containing medium. The effect of SDF-1α-LN crosstalk on NSCs migrated numbers was determined by using Transwell cell migration test, and the differentiation of NSCs was determined by immunofluorescence staining.
RESULTS
Primary NSCs were successfully isolated and cultivated, neurospheres formed at 3-5 days with typical NSCs morphology positively expressing Nestin which was the specific antigen of NSCs. Compared with PLL group, the number of NSCs migration in PLL+SDF-1α group showed no significant change (cells: 3.00±0.99 vs. 2.3±0.67, P > 0.05). Compared with LN group, the number of NSCs migration in LN+SDF-1α group was significantly increased (cells: 85.33±9.61 vs. 31.67±5.86, P < 0.05). Immunofluorescence staining showed that the differentiation rates of PLL and PLL+SDF-1α groups were almost zero, and some early differentiation neurons were detected in LN group with the differentiation rates of (12.50±2.56)%. The differentiation of early neuronal cells was significantly increased after SDF-1α-LN crosstalk, the neuronal differentiation rate was (21.40±3.41)%, and it was significantly higher than that of LN group (P < 0.05).
CONCLUSIONS
SDF-1α crosstalk with LN in extracellular matrix can significantly and synergistically enhance the migration and differentiation of NSCs in vitro.
目的
探讨基质细胞衍生因子-1α（SDF-1α）与层粘连蛋白（LN）相互作用在神经干细胞（NSCs）迁移和分化中的作用。
方法
分离培养从14天孕龄的胎鼠中获取的原代NSCs，采用免疫荧光法鉴定其表型特征。收集第三代NSCs，分为四组：聚-L-赖氨酸（PLL）组、PLL+SDF-1α组、LN组和LN+SDF-1α组。将PLL组和LN组的NSCs接种到涂有0.1 mg/mL PLL或1 mg/mL LN的培养板中，将PLL+SDF-1α组和LN+SDF-1α组的NSCs接种到涂有PLL或LN且含有1 mg/mL SDF-1α的培养基的培养板中。采用Transwell细胞迁移试验检测SDF-1α-LN相互作用对NSCs迁移数量的影响，通过免疫荧光染色检测NSCs的分化情况。
结果
成功分离培养原代NSCs，3-5天形成神经球，具有典型的NSCs形态，阳性表达NSCs的特异性抗原巢蛋白。与PLL组相比，PLL+SDF-1α组NSCs迁移数量无明显变化（细胞数：3.00±0.99 vs. 2.3±0.67，P>0.05）。与LN组相比，LN+SDF-1α组NSCs迁移数量显著增加（细胞数：85.33±9.61 vs. 31.67±5.86，P<0.05）。免疫荧光染色显示，PLL组和PLL+SDF-1α组的分化率几乎为零，LN组检测到一些早期分化神经元，分化率为（12.50±2.56）%。SDF-1α-LN相互作用后早期神经元细胞分化显著增加，神经元分化率为（21.40±3.41）%，显著高于LN组（P<0.05）。
结论
细胞外基质中SDF-1α与LN相互作用可显著协同增强体外NSCs的迁移和分化。
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