Stromal cell derived factor-1α-laminin crosstalk induced neural stem cells migration and differentiation in vitro / 中华危重病急救医学

ABSTRACT

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).Conclusion SDF-1α crosstalk with LN in extracellular matrix can significantly and synergistically enhance the migration and differentiation of NSCs in vitro.