微载体旋转立体培养成人退变髓核细胞及鉴定  

Adult degenerative nucleus pulpous cells cultured by a microcarrier-based stirred culture system

作　　者：宁斌[1] 丁远景[1] 龚维明[1] 刘海飞 刘勇 王德春 胡有谷 

机构地区：[1]山东大学附属济南市中心医院脊柱外科,山东省济南市250013 [2]山东省创伤骨科研究所,山东省青岛市266001

出　　处：《中国组织工程研究》2013年第7期1168-1173,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(30672132).

摘　　要：背景:旋转生物反应器是目前应用较多的微载体培养系统。目的:应用微载体旋转立体培养方法对成人退变髓核细胞进行扩增及鉴定。方法:对手术切除的髓核组织进行原代培养,传代后进行单层培养和微载体旋转立体培养的对比,并行苏木精-伊红、Gimsa染色观察细胞形态,利用电镜观察其超微结构,利用MTT比色法检测细胞生长活性。结果与结论:倒置相差显微镜观察培养的细胞符合退变髓核细胞形态,透射电镜下可显示其退变改变。倒置相差显微镜和扫描电镜下观察,可见退变的髓核细胞呈立体状生长,并能够成功甩珠传代。细胞在对数生长期时,微载体培养的细胞的生长活性明显高于单层培养组的细胞。表明微载体旋转立体培养法可以较好的维持细胞表型,对对数生长期髓核细胞的细胞活性具有明显的刺激作用。BACKGROUND： Rotating bioreactor is a microcarrier-based stirred culture system that is currently used more often. OBJECTIVE： To culture the adult degenerative nucleus pulposus cells in vitro by a microcarrier-based stirred culture system, and then to amplify and identify the cells. METHODS： Nucleus pulposus specimens taken from patients with disc degenerative diseases were randomly divided into monolayer culture group and micro-carrier culture group. The cell morphology was observed by hematoxylin-eosin staining and Gimsa staining, the microstructure was observed under anelectron microscope, cell viability was detected by MTT method RESULTS AND CONCLUSION： The morphology of degenerative nucleus pulpous cells was identified by inverted phase contrast microscope, and the regression was showed by a transmission electron microscope. The microcarrier cultured cells were observed by an inverted phase contrast microscope and scanning electron microscope showing stereoscopic shape. In the logarithmic growth phase, the growth activity of microcarrier culture cells was significantly higher than that of the cells in monolayer culture. Microcarrier-based stirred culture system can stabilize the cell phenotype, and significantly stimulate the cellular activity of nucleus pulposus cells in the logarithmic growth phase.

关 键 词：组织构建 软骨组织构建 髓核细胞 退变 微载体 细胞培养 细胞鉴定 立体培养 单层培养 国 家自然科学基金 组织构建图片文章 

分 类 号：R318[医药卫生—生物医学工程]