Journal of Jilin University Medicine Edition ›› 2017, Vol. 43 ›› Issue (01): 186-189.doi: 10.13481/j.1671-587x.20170137

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Primary culture and identification of mouse kidney cells

GAO Xia1, LEI Xiaoyan1, LIU Shurao2, SUO Yanhong3, CAO Xiaofeng1, GAO Mingdong1   

  1. 1. Department of Pediatrics, People's Hospital of Gansu Province, Lanzhou 730000, China;
    2. Department of Postgraduate, Gansu University of Traditional Chinese Medicine, Lanzhou 730000, China;
    3. School of Postgraduate, Ningxia Medical University, Yinchuan 750001, China
  • Received:2016-05-31 Online:2017-01-28 Published:2017-02-08

Abstract:

Objective: To establish a good operability, simple and efficient mouse kidney cell separation and primary culture model, and to lay the foundation for further study.Methods: The murine kidney tissues were collected and the glomeruli with different size combination of screening were obtained. The isolated glomeruli were suspensioned in KI-3T3 medium, then subsided in the medium cell with rat tail collagen. The glomeruli were refreshed 4 d after cultivation. 7 d after cultivation, the glomeruli were resolved by trypsin and the podocyts were cultured. The podocytes were generated at 5-7 d. The podocytes were analyzed after 2-3 generations. The morphology of podocytes of kidney tissue of the mice was observed by inverted phase contract microscope.The expressions of nephrin,podocin and P-cadherin in the podocytes of kidney tissue of the mice were detected by PCR.Results: The podocytes began to remove from the planted glomeruli at the 3th day. After 7 d of generation, the cultured podocytes showed microvilli and process.The PCR results showed that the podocytes expressed nephrin,podocin, and P-cadherin.Conclusion: The difference sifting technology combined with enzyme digestion could successfully isolate and culture the C57/BL6J mouse podocytes in vitro.

Key words: kidney, podocyte, primary culture

CLC Number: 

  • Q813.11