Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (50): 9335-9338.doi: 10.3969/j.issn.1673-8225.2011.50.007

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Cultivation of mouse mammary epithelial cells by using improved collagenase/trypsin digestion method

Yi Qiong1, Wang Lu1, Li Yuan-fang2, Zhu Wei2   

  1. 1Animal Science College of Guizhou University, Guiyang  550025, Guizhou Province, China
    2Center of Biochemical Engineering in Guizhou Province, Guiyang  550025, Guizhou Province, China
  • Received:2011-06-28 Revised:2011-10-26 Online:2011-12-10 Published:2011-12-10
  • Contact: Wang Lu, Doctor, Professor, Master’s supervisor, Animal Science College of Guizhou University, Guiyang 550025, Guizhou Province, China wanglu7007@163.com
  • About author:Yi Qiong★, Studying for master’s degree, Animal Science College of Guizhou University, Guiyang 550025, Guizhou Province, China yiqiong19870328@126.com
  • Supported by:

    the National Nature Science Foundation of China, No. 31060347*; Innovation Foundation for Graduate in Guizhou University, No. Xiaoyannong 2010020*

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Abstract:

BACKGROUND: The mixture of collagenase A and trypsin that used to isolate the mammary epithelial cells is complex and strict to operate.
OBJECTIVE: To clarify whether mammary epithelial cells can be successively cultured with improved collagenase/trypsin digestion method in vitro.
METHODS: The mixture of typeⅠand Ⅱcollagenase and trypsin (1:1:1) was used to digest breast tissue, which was shredded directly with ophthalmic scissors. The organoids were obtained by 37 ℃ oscillation extraction and the differential adhesion method was used to remove the fibroblasts. Then the organoids were inoculated in the plastic dish.
RESULTS AND CONCLUSION: Inverted microscope observation showed that epithelial organoids could be mostly acquired and had attached to the plastic dish after 12 hours, and then cells were spreading out from them and forming confluent monolayer of cobblestone after cultured for 72 hours. In addition, the tissue-specific expression of cytokeratin 18 in mammary epithelial cells was identified by cytokeratin immunohistochemistry. It indicated that plenty of pure mammary epithelial cells could be harvested by using improved collagenase/trypsin digestion.

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