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Published on: August 23, 2016
[Stromal cell derived factor-1α-laminin crosstalk induced neural stem cells migration and differentiation in vitro]
Jianqi Zhang1, Xiaohong Li, Sai Zhang
1Department of Cardiovascular Medicine, Affiliated Hospital of Logistics College of Armed Police Force, Tianjin 300162, China. Corresponding author: Zhang Jianqi,
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.
