兔髂骨生长板细胞的培养及体内成软骨作用的研究  

The Culture of the Iliac Growth-plate Chondrocytes and the Study on Its Cartilage-formation Role in Vivo

作　　者：靳小兵[1] 罗卓荆[1] 杨柳[1] 兰丽锋[1] 

机构地区：[1]第四军医大学西京医院全军骨科研究所,西安市710032

出　　处：《骨与关节损伤杂志》2003年第8期546-548,共3页The Journal of Bone and Joint Injury

基　　金：国家教育部高等院校骨干教师资助计划项目

摘　　要：目的 建立大量生产髂骨生长板软骨细胞的培养及保存方法,观察髂骨生长板细胞在裸鼠体内的成软骨作用,为生长板组织工程提供种子细胞。方法 取二周龄兔髂骨生长板软骨,经机械剪切和化学消化作用后,接种、培养及液氮保存,通过显微镜观察其生物学表现,并通过甲苯胺兰、碱性磷酸酶染色及Ⅱ型胶原染色对其生物学特性进行签定。将生长良好的第二代细胞注入裸鼠背部,于第四周取材,进行组织学观察。结果 细胞可在体外大量增殖并在六代内无明显反分化,但缺乏向肥大细胞进一步分化的能力。冷冻复苏细胞存活率为92％。注射的细胞悬液可在裸鼠体内形成软骨组织,并进一步向肥大细胞转化。结论 成功建立了髂骨生长板细胞的培养及冻存方法,证明髂骨生长板细胞可在体内进一步形成软骨组织并向肥大期细胞转化。Objective To establish a kind of iliac growth-plate chondrocyte culture and storage method so that we can obtain the seed cells in large quantity for the usage of engineered growth plate tissue. To observe the cartilage formation of the cells in vivo. Methods The cells were harvested from the iliac growth-plate cartilage of the two-week old rabbit and cultured. Their biological phenomenons were observed and their biological characteristics were identified by using toluidine blue, alkaline phosphatase staining and immunohistochemistry staining for type II collagen. The second generation cells were harvested and injected into the dorsum of nude mice. Specimens were harvested 4 weeks later for histological examination. Results The cells could proliferate in vitro and have no apparent de-differentiation within the sixth generation, but they could not differentiate into the hypertrophic cells. The cells revived from being frozen ( - 196℃ ) was 92% . The cells injected into the body of the nude mice could form the cartilage-like tissue. The iliac growth-plate chondrocytes can differentiate into hypertrophic cells in vivo. Conclusion We have established a method of culture and storage of iliac growth-plate chondrocytes and cartilage-formation of the cells in vivo was verified. The cells can differentiate into hypertrophic chondrocytes in vivo.

关 键 词：髂骨生长板 细胞培养 体内成软骨作用 组织工程 

分 类 号：R329-33[医药卫生—人体解剖和组织胚胎学]