Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (11): 1973-1076.doi: 10.3969/j.issn.1673-8225.2011.11.018

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Culture and identification of rat annulus fibrosus cells

Zhang Kun-mu1, 2, Chen Shao-qing1, 3, Song Hong-mei2, 4, He Yan5, Liao Jun1, 3, Hong Yu1, 4, Wang Shi-zhong1, 2, 3   

  1. 1Institute of Rehabilitative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou  350003, Fujian Province, China
    2Second People’s Hospital, Fujian University of Traditional Chinese Medicine, Fuzhou  350003, Fujian Province, China
    3 TCM Rehabilitating Technique Level 3 Laboratory of State Administration of Traditional Chinese Medicine, Fuzhou  350003, Fujian Province, China
    4Fujian Movement Function Rehabilitation Key Laboratory, Fuzhou  350003, Fujian Province, China
    5Laboratory of Biochemistry and Molecular Biology, Fujian Medical University, Fuzhou  350000, Fujian Province, China
  • Received:2010-10-16 Revised:2011-01-12 Online:2011-03-12 Published:2011-03-12
  • Contact: Wang Shi-zhong, Doctor, Professor, Chief physician, Doctoral supervisor, Institute of Rehabilitative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350008, Fujian Province, China; Second People’s Hospital of Fujian University of Traditional Chinese Medicine, Fuzhou 350003, Fujian Province, China; TCM Rehabilitating Technique Level 3Laboratory of State Administration of Traditional Chinese Medicine, Fuzhou 350003, Fujian Province, China fujiantuina@126.com
  • About author:Zhang Kun-mu★, Studying for master’s degree, Institute of Rehabilitative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350008, Fujian Province, China; Second People’s Hospital of Fujian University of Traditional Chinese Medicine, Fuzhou 350003, Institute of Rehabilitative medicine Fujian Province, China fujiantuina@126.com
  • Supported by:

    the National Natural Science Foundation of China, No. C190203*; the Science Research Foundation of Ministry of Health, No. WKJ2008-2-48*; International Cooperation Project of Science and Technology Commission of Fujian Province, No. 2009I0010*; Development Fund of Chen Ke-ji Traditional and Western Medicine, No. Ckj2008012*

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Abstract:

BACKGROUND: There is not a criterion for culture of annulus fibrosus cells, thus, there is deficiency in operability and repetitiveness.
OBJECTIVE: To establish the annulus cell lines and to observe their continuouschanges, in order to provide the methods and cell model for studying molecular biology in tissue engineering field.
METHODS: The annulus fibrosus cells were isolated with enzyme digestion and grew as monolayer. DMEM containing 15% fetal calf serum was used in the primary culture, which was replaced by DMEM containing 10% fetal calf serum after cell passage. Morphology, structure and biological characteristics of cells were detected by an inverted phase contrast microscope. The phenotype was identified with the toluiding blue staining and immunocytochemistry.
RESULTS AND CONCLUSION: The disc fibrinous ring cells were cultured in monolayer with the round or fusiformis morphology. The primary cells grew more slowly than the subculture cells. The cells displayed intense toluiding blue metachromasia; the immunohistochemical test revealed that the positive expression of type Ⅰ and Ⅱ collagen on the cells. The experiment successfully culture annulus fibrosus cells in vitro. 

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