牛髓核软骨样细胞原代培养及异种移植组织相容性观察  

Primary culture of bovine chondrocyte-like nucleus pulposus cells and cell histocompatibilty in xenotransplantation

作　　者：侯晓飞[1] 李浩鹏[1] 杨保辉[1] 

机构地区：[1]西安交通大学医学院第二附属医院骨二科,陕西省西安市710004

出　　处：《中国组织工程研究与临床康复》2011年第44期8205-8208,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:自体或同种异体细胞移植进行椎间盘再生已成为临床研究的热点。目的:探索牛椎间盘髓核软骨样细胞原代提取条件及培养方法,观察其植入兔椎间盘中组织相容性。方法:提取新鲜的牛尾椎间盘中的髓核组织,分离并体外培养牛髓核软骨样细胞。经腹后外侧途径采用微量加样针将扩增过的牛髓核软骨样细胞注入新西兰大白兔的椎间盘中。分别在移植后的2,4,6及8周收集接受牛髓核软骨样细胞移植的兔椎间盘,以冰冻免疫组织化学法检测CD5或CD4细胞表面抗原的表达及B或T淋巴细胞的浸润,判断是否发生免疫排异反应。结果与结论:实验分离出了活牛软骨样细胞,在体外进行了传代扩增。在移植后2,4,6及8周对L3/4,L4/5,L5/6椎间盘及腹腔淋巴结行抗兔CD4、CD5染色后未发现阳性表达,椎间盘局部无B或T淋巴细胞的浸润。移植入兔椎间盘8周,经冰冻免疫组织化学检测未发现免疫排异反应。说明原代培养的牛椎间盘髓核软骨样细胞组织相容性较好。BACKGROUND： Autologous or allogenic cell transplantation for regeneration of intervertebral discs has become a hot spot in clinical study. OBJECTIVE： To investigate primary culture method of bovine chondrocyte-like nucleus pulposus cells and observe cell histocompatibility after transplantation into rabbit intervertebral discs. METHODS： Fresh bovine caudal nuclesus pulposus was collected. Bovine chondrocyte-like nucleus pulposus cells were isolated, sequentially digested by pronase E and collagenase Ⅱ and cultured in DMEM/F12 medium and 10 %fetal bovine serum. The expanded cells were injected into rabbit intervertebral discs through the use of microinjector via the posterolateral approach. At 2, 4, 6 and 8 weeks after transplantation, rabbit intervertebral discs in which bovine caudal nuclesus pulposus cells were transplanted. CD5 or CD4 cell surface antigen expression and B or T lymphocyte infiltration were detected using immunohistochemistry. RESULTS AND CONCLUSION： Bovine chondrocyte-like cells were isolated successfully. Their number was expanded in vitro. At 2, 4, 6 and 8 weeks after transplantation, no anti-rabbit CD4 and CD5 immunoactive expression or C or T lymphocyte infiltration was observed in any treated intervertebral discs. At 8 weeks after transplantation, no immunological rejection was observed, as shown by immunohistochemistry. These findings suggest that primary cultured bovine chondrocyte-like nuclesus pulposus shows good histocompatibility.

关 键 词：椎间盘退变 细胞原代培养 异种移植 组织相容性 组织工程 

分 类 号：R617[医药卫生—外科学]