Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (46): 8593-8598.doi: 10.3969/j.issn.2095-4344.2012.46.009

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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:2013-03-16
  • 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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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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