成人退变髓核细胞体外培养和细胞周期测定  

In vitro culture and cell cycle detection of the adult degenerated nucleus pulposus

作　　者：邱匀峰 吴小涛[2] 赵梓汝[1] 王运涛[2] 

机构地区：[1]东南大学临床医学院,南京210009 [2]东南大学附属中大医院骨科,南京210009

出　　处：《中国矫形外科杂志》2009年第1期52-54,共3页Orthopedic Journal of China

基　　金：东南大学国家自然基金预研项目资助(编号:XJ0690256)

摘　　要：[目的]建立成人退变髓核细胞的单层培养模型并观察其形态;测定其细胞周期,探讨传代后髓核细胞生长不佳的原因。[方法]取24例椎间盘突出症患者手术切除的椎间盘组织,分离出髓核组织,胰酶和胶原酶消化,DMEM/12培养基培养。倒置相差显微镜观察其形态学特征,流式细胞分析仪检测原代和P2细胞周期。[结果](1)原代髓核细胞生长良好,7 d后贴壁生长的细胞可达90%。(2)原代细胞凋亡率为(38.10±11.7)%,P2代凋亡率为(44.74±17.6)%,原代S期细胞(7.88±2.1)%,P2 S期细胞(2.76±0.7)%。[结论]传代后的髓核细胞凋亡率增高,S期细胞减少明显。[ Objective] To establish an in vitro two -dimensional culture model of degenerated nucleus pulposus and detect samples＇ cell cycle by flow cytometry to study why nucleus pulposus cells don＇t grow well after passage. [ Method] Nucleus pulposus tissues taken from protruded discs of 24 patients were treated by Trypsin and collagenase after surgical procedures, and then the cells were cultured in DMEM/F12 medium. Cell morphology was observed by an invert microscope and cell cycle of the primary and P2 cells were detected by flow cytometry after proliferation in monolayer culture. [ Result ] 1. Primary culture cells of nucleus pulposus grew well in the medium, and 90% cells adhere to layer after about 7d. 2. The rate of apoptosis of NP： primary （38.10±11.7）%, P2 （44.74±17.6）%. The rate ors period ： primary （7.88±2.1%）, P2 （2.76 ±0.7）%. [ Conclusion] When going down to posterity the cells＇ apoptosis rate grows while S period cells decrease.

关 键 词：椎间盘 髓核细胞 细胞周期 凋亡 

分 类 号：R681.53[医药卫生—骨科学]