成骨细胞体外培养及异体移植免疫学检测

Osteoblast Primary Culture and Immunological Research of Allotransplantation

目的 :成骨细胞原代培养 ,检测其异体移植免疫反应情况 ,探讨其免疫学机制 ,为骨细胞移植及组织工程骨的构建寻找细胞来源。方法 :用酶消化 -组织块联合培养法行细胞原代培养 ,传代后行同胎及同种异系腹直肌袋内移植 ,于不同时间点行免疫学及形态学检测。结果 :原代接种后 1周细胞成单层 ,碱性磷酸酶染色阳性 ;2组分别于移植后2周及 4周检测到特异性抗体 ,同种异系组在移植后 2周检测到细胞介导的细胞毒反应 ,至 4周达高峰。结论

Objective: To culture osteoblast and detect the immunoreaction after allotransplantation,discuss the potential immunological mechanism and find the reliable cell source for osteocyte transplantation or tissue engineered bone reconstruction.Methods:Osteoblast was isolated by bony digestion/elements in culturing,then the cells of same venter and allostrip were transplanted into the rectus abdominus muscle of the each rabbit.Immunological tests and histological observation were performed 1 week?2 week?4 week and 8 week following transplantation.Results:Cells reached confluence within 1 week and positive result was detected in ALP staining.A specific antibody reaction were detected in both groups two weeks and four weeks after transplantation..Cell mediated cytotoxicity was detected in allostrip group two weeks after transplantation,and reached the peak value four weeks later.Conclusion: Same venter and allostrip osteoblast can be the reliable cell sources for osteocyte transplantation or tissue engineering bone reconstruction.