兔骨髓成骨细胞同种异体移植免疫反应的初步观察

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

A Preliminary Study on the Immune Responses during Allografting of Rabbit Bone Marrow Stromal Cells

To search for the ability of rabbit bone marrow stromal cells to induce the differentiation into osteoblasts and the possibility of allografting osteoblasts,the bone marrow cells were obtained fron tibia of newborne rabbits and cultivated in vitro in conditional DMEM medium with 10% of fetal bovine serum After 5 generations of passages,the cells were examined with Wrights staining,alkaline phosphatase (ALP) staining,and red alizarin staining Several pieces of the gelfoam absorbed revived cells were transplanted subcutaneously and intramuscularly Speciemens were observed after 2,4,8 and 12 weeks of operation,and the ability to form osteoblasts as well as the allograft rejection reaction were analyzed The results showed that the bone marrow osteoblasts of the in vitro induced cultures were a comparatively pure cell lines with spindle shape and with several protuberances,and to be positive in the ALP staining After cultivation in vitro for 40 days,many mineralized nodes were found around the cultured cells Majority of the transplanted osteoblasts were survived and after 4 weeks the transplanted cells began to produce osteoid matrix with irregular mineralized bone tissue Around the transplanted cells,there was only small amount of lymphocyte or eosinphil infiltrations It concludes that BMSC have significantly osteogenic potential and the transplanted cells still perserve their fundamental biological functions with slight degree of rejection reactions These results suggest it is possible for the performance of cell allografting