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以观察新生兔骨髓诱导分化为成骨细胞的能力 ,探讨同种异体成骨细胞移植的可行性 ,为进一步的研究奠定基础。取新生新西兰大白兔胫骨骨髓 ,分离后加入条件培养液体外培养。传 5代后对细胞进行形态学、碱性磷酸酶 (ALP )染色及体外矿化能力的检测。将冻存复苏的细胞以明胶海绵吸附 ,植入异体成年兔皮下、肌肉内 ,第 2、 4、 8、 12周时取材观察 ,分析其成骨能力及免疫排斥反应情况。结果显示体外诱导培养的骨髓成骨细胞是一较纯的细胞系 ,以带突起的梭形细胞为主 ,ALP染色阳性 ,连续培养 4 0d可见矿化结节形成。异体植入的成骨细胞大部分存活 ,4周后开始有类骨基质形成 ,并可见不规则的矿化骨组织 ,植入细胞周围仅见少量的淋巴细胞和嗜酸粒细胞浸润。骨髓基质细胞具有多向分化的潜力 ,异体植入的细胞仍保持基本的生物学功能 ,免疫排斥反应比较轻微 ,提示细胞异体移植是可行的
To observe the newborn rabbit bone marrow-derived osteoblast differentiation and explore the feasibility of allogeneic osteoblast transplantation for further research and lay the foundation. New Zealand white rabbits were taken from the tibia bone marrow, separated and cultured in vitro conditioned medium. After passage 5, the cells were examined for morphology, alkaline phosphatase (ALP) staining and mineralization in vitro. The frozen-thawed cells were adsorbed with gelatin sponge and implanted subcutaneously and intramuscularly in allogeneic adult rabbits. The cells were harvested at the 2nd, 4th, 8th and 12th weeks after operation. The osteogenic capacity and immune rejection were analyzed. The results showed that the cultured osteoblasts in vitro were a pure cell line, mainly with protruding spindle cells and positive ALP staining. The mineralized nodules were seen after continuous culture for 40 days. Most of allogeneic osteoblasts survived. After 4 weeks, osteoblasts began to form, and irregular mineralized bone tissues were seen. Only a small amount of lymphocytes and eosinophils infiltrated around the implanted cells. Bone marrow stromal cells have the potential of multi-directional differentiation, allogeneic cells still maintain the basic biological function, immune rejection is relatively mild, suggesting that allogeneic transplantation is feasible