摘要
目的通过对成人退变椎间盘髓核细胞的体外培养和形态学观察,进一步研究细胞因素在椎间盘退变中的作用机制。方法取5例患椎间盘突出症并行椎间盘摘除手术的成人髓核,分离后在培养基中进行髓核细胞培养,细胞染色、逆转录聚合酶链式反应、免疫荧光检测细胞Ⅰ、Ⅱ型胶原的表达。结果椎间盘髓核细胞可在体外培养,30d后方可进行传代,最佳的培养条件为胎牛血清/培养基体积分数为10%～20%,pH值7.0。髓核细胞中出现Ⅰ型胶原,并具有较高的表达,Ⅱ型胶原表达微弱。结论成人退变髓核细胞体外培养时间较长,细胞增殖能力低下,特定培养条件的摸索是成功与否的关键。
Objective This study is to explore the disc degeneration influenced by the cell effects by cells culture of adult degenerative lumbar nucleus pulposus disc.Methods Five discs were obtained from five adults with disc herniation disease by perations.They were dissected and were digested with trypsin and collagenase.The isolated cells were cultured in DMEM/12.The cultured cells were identified with morphological observation and semiquantitative RT-PCR etc.Results The NPCs can be cultured in vitro,After 30 days it just can be subcultured.The proper culture conditions include bulk percentum of calf cerum and PH value of DMEM.The cultured NPCs show high expressions of collagen Ⅰand low expression of collagen Ⅱ.Conclusion NPCs culture in vivo the culture need longer time,also the proliferous ability is lower.where the shoe pinches is to explore the cell culture conditions.
出处
《实用骨科杂志》
2011年第10期902-904,I0001,共4页
Journal of Practical Orthopaedics
基金
国家自然科学基金课题资助项目(30400453)
关键词
椎间盘
退变
髓核细胞
细胞培养
intervertebral disc
degenerative
nucleus pulposus
cell culture
