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背景:目前尚未见成熟的成年人退行性变椎间盘的细胞培养模型报道,而此模型为建立椎间盘组织工程的细胞学基础。目的:建立成人椎间盘细胞培养模型,奠定椎间盘组织工程研究的细胞学基础。设计及地点:对比观察的细胞学实验,在山东省创伤骨科研究所完成。材料:标本来源于青岛大学附属医院骨科5例确诊为退行性椎间盘病变患者。L2~3椎间盘1例,L4~5椎间盘3例,L5~S1椎间盘1例。方法:对5例取自腰椎间盘突出症退变椎间盘的手术标本,采用系列酶消化法细胞培养和组织块细胞培养两种方法,应用HAMF12培养基加体积分数为10%和20%的胎牛血清分别进行了细胞培养,均获得了传代,并对培养细胞进行了光镜和电镜的形态学观察。主要观察指标:①细胞生长状况。②光镜下细胞形态观察。③透射电镜超微结构观察。结果:酶消化法和组织块法均能获得大量的退变椎间盘细胞,并成功地进行细胞传代。培养细胞的分泌物质可能对细胞的生长起一定的限制作用,去除该物质后细胞可迅速增生。但在细胞生长及增殖过程中,体积分数为20%胎牛血清下细胞增殖速度明显高于体积分数为10%胎牛血清时的速度。全部退变椎间盘标本中均发现了脊索细胞。结论:成人退变椎间盘可以用来获得椎间盘组织工程所需的细胞。
BACKGROUND: There are no cell culture models reported in mature adult degenerative intervertebral discs, and this model is a cytological basis for the establishment of intervertebral disc tissue engineering. OBJECTIVE: To establish an adult model of intervertebral disc cell culture and establish the cytological basis for the tissue engineering research of disc tissue. Design and Location: Comparative observation of cytology experiments in Shandong Province orthopedic Institute completed. Materials: Specimens from the Department of Orthopedics, Affiliated Hospital of Qingdao University, 5 patients diagnosed as degenerative disc disease patients. 1 case of L2 ~ 3 disc, 3 cases of L4 ~ 5 disc, and 1 case of L5 ~ S1 disc. Methods: Five cases of degenerated intervertebral disc herniated by lumbar disc herniation were treated with series of enzyme digestion and tissue cell culture. HAMF12 medium supplemented with 10% and 20% fetal calf Serum cells were cultured, were obtained passaged, and the cultured cells were observed by light microscopy and electron microscopy. MAIN OUTCOME MEASURES: ① cell growth status. ② light microscope observation of cell morphology. ③ transmission electron microscopy ultrastructural observation. Results: A large number of degenerated intervertebral disc cells were obtained by enzyme digestion and tissue block method, and the cells were successfully passaged. Secretion of cultured cells may play a limiting role in the growth of cells, cells can rapidly proliferate after removal of the substance. However, in the process of cell growth and proliferation, the rate of cell proliferation under 20% fetal bovine serum was significantly higher than that at 10% fetal bovine serum. Chordate cells were found in all degenerated disc specimens. CONCLUSIONS: Adult degenerative discs can be used to obtain the cells needed for disc tissue engineering.