Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (24): 4398-4401.doi: 10.3969/j.issn.1673-8225.2010.24.007

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Primary culture of fibroblasts from cochlear duct membrane of neonatal C57 mouse using enzyme digestion and differential adhesion methods  

Wang Ting-kuo, Sun Hong   

  1. Department of Otorhinolaryngology-Head & Neck Surgery, Third Xiangya Hospital, Central South University, Changsha  410013, Hunan Province, China
  • Online:2010-06-11 Published:2010-06-11
  • Contact: Sun Hong, Doctor, Professor, Department of Otorhinolaryngology-Head & Neck Surgery, Third Xiangya Hospital, Central South University, Changsha 410013, Hunan Province, China shjhaj@vip163.com
  • About author:Wang Ting-kuo☆, Studying for doctorate, Attending physician, Department of Otorhinolaryngology-Head & Neck Surgery, Third Xiangya Hospital, Central South University, Changsha 410013, Hunan Province, China wtk711112@163.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30371531*; the Natural Science Foundation of Hunan Province, No. 02JJY2050*; the Scientific Research Fund of Health Department of Hunan Province, No. B2006-063*; the Science and Technology Planning Project of Hunan Province, No. 2007SK2001*

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Abstract:

BACKGROUND: There has been no specific method for culturing fibroblasts from cochlear duct membranes so far.
OBJECTIVE: To provide good cell model in vitro via culturing fibroblasts from lateral wall of cochlear duct membrane of neonatal C57 mouse by enzyme digestion and differential adhesion methods and to identify its biological features.
METHODS: Tissues from lateral wall of cochlear duct membrane were acquired from neonatal C57 mouse using operating microscope. Fibroblasts from lateral wall of cochlear duct membrane were cultured by trypsin digestion combined with differential adhesion methods. The growth condition was observed by inverted phase contrast microscope and hematoxylin-eosin staining, and cell growth curve was drawn, then the immunocytochemistry was employed to classify cell types.
RESULTS AND CONCLUSION: After passage and purification, the shape of fibroblasts from lateral wall of cochlear duct membrane was fusiform and triangle, with “S” type cell growth curve. Immunocytochemiscal detection showed that vimentin could be detected in cultured cells, and brown cytoplasmic pigment could be seen. The results demonstrated that fibroblasts can be cultured form lateral wall of cochlear duct membrane of mice. 

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