| Objective: To observe changes of injured spinal cord after transplanted human hair keratin (HHK) combined with neural stem cells (NSCs), and evaluate motor function of lower limbs after operation. To investigate the feasibility of repairing spinal cord injury by tissue engineering, so as to provide a new consideration of NSCs transplantation in treating nervous system disease or injury.Methods: 1.Isolation, cultivation and characterization of NSCs. The cells were derived from embryonic rat (E14-16) cerebral cortex, planted in DMEM/F12 serum-free medium with epidermal growth factor, basic fibroblast growth factor, leukaemia inhibitory factor and N2. Cells in passage 5 to 10 were marked with 5-bromodeoxyuridine (BrdU) after they were identified by immunofluorescence technique.2.Empirical study on co-transplantation of HHK and NSCs in rats with spinal cord injury. 80 adult female SD rats randomly allocated into 4 groups according to factorial design: SCI group, HHK implantation group, NSCs transplantion group and co-transplantation group. According to BBB locomotor rating scale, we prepared moderate SCI model (T10) by NYU method. HHK and NSCs were implanted into injured region immediately after SCI models were established. We observed the changes of behavior after operation, evaluted motor funtion of lower limbs by BBB scoring and tiltboard experiment.We also investigated the proliferation, differentiation and migration of NSCs in injured spinal cord after transplantation, and the integration of NSCs, HHK in host spinal cord.Results: 1.Isolation, cultivation and characterization of NSCs: The cultured cellsformed suspended neurosphere, showing strong capability of proliferation and self-renewal. The cells were Nestin positive when identified by laser scanning confocal microscopy, so we make sure that the cells were NSCs. 2.Histological changes in injured region: 1st week: Spinal cord edema and gray...
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