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Objective: (1) separation, culture and identification of SD rat BMSCs induced differentiation of Nestin positive the BMSCs and Ang-1 gene was transfected into cells by molecular biology techniques; (2) Construction of SD rats T10 spinal cord clamp injury model to assess its feasibility; (3) Ang-1 gene-modified the Nestin positive BMSCs local injection treatment for spinal cord injury detected Nestin-positive BMSCs can differentiate into neurons, detection of Ang-1 transplanted into the body of the the Nestin positive BMSCs newborn and remaining after the injury neurons the protective effect. Methods: (1) the use of whole bone marrow differential adhesion isolated and cultured the SD rats hind limbs within the femur, tibia bone marrow BMSCs were isolated and cultured. Traits of cell growth was observed under a microscope; flow cytometry cell surface marker objects CD29, CD34, CD45 and CD90 expression; osteoblast-like cells into fat-like cells into neural-like cells demonstrate that their multipotent differentiation potential; BMSCs induced to differentiate into Nestin positive the BMSCs and Ang-1 gene was transfected into adenovirus vector Nestin positive BMSCs Weston blot detection after transfection, cells whether expression of Ang-1. (2) Construction of SD rats T10 spinal cord clamp injury model, modeling after 1 week evaluation of motor function, nerve electrophysiological testing and histopathology testing the feasibility of the method detection model; (3) modeling 1 week after Nestin-positive rats were randomized implementation of local injection of transplanted cells were treated in the subsequent 1 to 4 weeks motor function assessment, neurophysiological testing and spinal cord tissue immunohistochemistry method to detect genetically modified Ang-1 The BMSCs spinal cord injury. Results: (1) differential through the whole bone marrow adherent proliferation of BMSCs were isolated and cultured speed is fast, amplification ability, subculture 3rd generation cells have shown a higher purity; cell surface markers by flow cytometry, cells expressing CD29 and CD90, the positive rate of more than 99%; cells can differentiate into bone cells in vitro by inducing differentiation into fat cells into neuron-like cells; Weston blot detection after transfection Nestin positive BMSCs Ang-1 strip. (2) modeling after 1 week, rats significantly the paraplegia (BBB lt; 4 points), the the neurophysiological test can not lead CSEP waveform, spinal cord tissue inflammatory cell infiltration, vacuolar degeneration of the spinal cord. (3) after transplantation 1-4 weeks A group hindlimb no significant changes in nerve function, B, C group rats were varying degrees of recovery, group C than in group B was significantly (P lt; 0.05); neurophysiological detection, C group were significantly better than in group B (P lt; 0.05); spinal cord injury after immunohistochemical staining, B, C in both groups express BrdU, Nestin, NSE positive cells gradually decreased with time, 4 weeks after transplantation group C remains a certain amount of positive cells, and significantly more than A, B two groups (P lt; 0.05). Group C were also observed BrdU-positive cells gathered around the lumen. Conclusion: (1) through the whole bone marrow differential method to obtain higher purity of BMSCs were isolated and cultured adherent cells pluripotent, after neural induction, cells are able to differentiate into Nestin positive of BMSCs by molecular biology techniques, Ang 1 gene was transfected into Nestin-positive BMSCs to be successful. (2) successfully constructed clamp spinal cord injury model in SD rats T10. (3) the Nestin positive after Ang-1 gene-modified BMSCs transplanted into the body to further differentiate into mature neurons, the secretion of Ang-1 Nestin-positive BMSCs, newborn and residual nerve cell has a protective effect, spinal cord injury repair.
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