Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (37): 6846-6850.doi: 10.3969/j.issn.1673-8225.2011.37.002

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Effects of basic fibroblast growth factor on cartilaginous tissue formation from epiphyseal plate chondrocytes cultured in centrifuge tube

Li Wen-chao1, Xu Rui-jiang1, Huang Jing-xiang2 , Zhang Li2, Nie Shao-bo1   

  1. 1Department of Pediatric Surgery, People’s Liberation Army General Hospital, Beijing 100853, China; 2Institute of Orthopedics, People’s Liberation Army General Hospital, Beijing 100853, China
  • Received:2011-03-09 Revised:2011-06-16 Online:2011-09-10 Published:2011-09-10
  • Contact: Xu Rui-jiang, Chief physician, Professor, Doctoral supervisor, Department of Pediatric Surgery, People’s Liberation Army General Hospital, Beijing 100853, China xurj301@yahoo. com.cn
  • About author:Li Wen-chao☆, Studying for doctorate, Department of Pediatric Surgery, People’s Liberation Army General Hospital, Beijing 100853, China liwenchao0311@163.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30772276*; Science and Technology Innovation Program of People’s Liberation Army General Hospital, No.062S02*

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Abstract:

BACKGROUND: There have been many reports regarding epiphyseal plate cells cultured in the centrifuge tube technique.
OBJECTIVE: To observe the effects of basic fibroblast growth factor (bFGF) on cartilaginous tissue formation from epiphyseal plate chondrocytes cultured in centrifuge tube.
METHODS: Chondrocytes were isolated from 3-week-old New Zealand rabbits using the method of tissue-yarn. A total of 5×106 chondrocytes obtained by centrifugation were cultured in a plastic centrifuge tube (15 mL) with the DMEM culture fluid including
10 μg/L bFGF for 4 weeks continuously. Cell morphology was investigated with light and inverted microscope, and the histological structure of new cartilaginous tissue was observed by histological staining.
RESULTS AND CONCLUSIONS: Cartilaginous tissue regenerated from growth plate chondrocytes cultured in centrifuge tube with bFGF in the DMEM culture medium. The periphery of the cartilaginous tissue was resembled with the germinal layer of growth plate, consisting of several cellular layers. The chondrocytes in the centre of tissue grew well, and several were differentiating to the mast chondrocytes. The cartilaginous tissue appeared strongly positive for safranine "O" and toluidine blue staining, which showed that the chondrocytes could synthesize the proteoglycans. Type II collagen immunohistochemistry appeared strongly metachromatic. bFGF could promote the formation of cartilaginous tissue, rich in proteoglycans and type II collagen, from epiphyseal plate cells cultured in centrifuge tune.

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