中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (46): 8593-8598.doi: 10.3969/j.issn.2095-4344.2012.46.009

• 软骨组织构建 cartilage tissue construction • 上一篇    下一篇

犬关节软骨细胞的体外分离与培养

王朝强1, 2，田丰德2，谢 辉2，王本杰2，刘保一2，赵德伟2   

  1. 1福建医科大学附属闽东医院，福建省宁德市 355000
    2大连大学，辽宁省大连市 116001
  • 收稿日期:2012-03-18 修回日期:2012-04-16 出版日期:2012-11-11 发布日期:2012-11-11
  • 通讯作者: 赵德伟，博士，主任医师，教授，大连大学，辽宁省大连市 116001
  • 作者简介:王朝强★，男，1986年生，福建省福清市人，汉族，2012年大连大学毕业，硕士，主要从事骨关节软骨损伤方面研究。 chaos911@163.com 并列第一作者：田丰德★，男，1977年生，辽宁省沈阳市人，汉族，2007年遵义医学院毕业，硕士，主要从事骨关节损伤方面研究。 tianfengde@163.com

Isolation and culture of canine articular chondrocytes in vitro

Wang Chao-qiang1, 2, Tian Feng-de2, Xie Hui2, Wang Ben-jie2, Liu Bao-yi2, Zhao De-wei2   

  1. 1Mindong Hospital, Fujian Medical University, Ningde 355000, Fujian Province, China
    2Dalian University, Dalian 116001, Liaoning Province, China
  • Received:2012-03-18 Revised:2012-04-16 Online:2012-11-11 Published:2012-11-11
  • Contact: Zhao De-wei, Doctor, Chief physician, Professor, Dalian University, Dalian 116001, Liaoning Province, China
  • About author:Wang Chao-qiang★, Master, Mindong Hospital, Fujian Medical University, Ningde 355000, Fujian Province, China; Dalian University, Dalian 116001, Liaoning Province, China chaos911@163.com Tian Feng-de, Master, Dalian University, Dalian 116001, Liaoning Province, China tianfengde@163.com Wang Chao-qiang and Tian Feng-de contributed equally to this study.

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496

被引次数

6

摘要：

背景：自1967年Manning等提出的通过胰蛋白酶和细菌胶原酶联合消化法分离软骨细胞以来，软骨细胞的体外分离培养研究较多，但目前尚无统一标准。
目的：研究犬关节软骨细胞在体外分离以及培养的条件，寻求体外扩增软骨细胞较简便、可行、高效的实验方法。
方法：取3周龄幼犬关节软骨组织，应用胰酶、胶原酶制定8种消化方法获取软骨细胞，对比不同方法所获取细胞数量及细胞成活率，分离细胞进行原代及传代培养，观察各代软骨细胞形态。
结果与结论：在体外分离犬关节软骨细胞的各种方法中，单纯应用Ⅱ型胶原酶消化法获得的软骨细胞数最多，细胞成活率最高。软骨细胞可以通过体外培养获得扩增，并且维持良好的细胞形态及表型，但仅限于5代以内。

关键词: 软骨细胞, 细胞培养, 免疫组化, 组织工程, Ⅱ型胶原酶, 胰酶

Abstract:

BACKGROUND: Since Manning et al reported to isolate chondrocytes using trypsin and bacterial collagenase digestion in 1967, in vitro isolation and culture of chondrocytes has been widely studied, but there is still no unified standard.
OBJECTIVE: To study the conditions for in vitro culture and isolation of chondrocytes and to explore a simple, feasible, and efficient experimental method to isolate, culture and proliferate canine chondrocytes in vitro.
METHODS: The articular cartilage of 3-week-old puppies were isolated and digested with trypsin and type Ⅱ collagenase for harvesting chondrocytes in different conditions. We compared the number of cells cell survival rate obtained by different methods. Isolated cells were passaged and morphology of chondrocytes was observed.
RESULTS AND CONCLUSION: The number and survival rate of chondrocytes was highest by using simple type Ⅱ collagenase digestion. Chondrocytes can be amplified through in vitro culture and maintain good cell morphology and phenotype, but can only be passaged less than five passages.

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