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Chinese Journal of Clinicians(Electronic Edition) ›› 2017, Vol. 11 ›› Issue (14): 2033-2037. doi: 10.3877/cma.j.issn.1674-0785.2017.14.005

Special Issue:

• Basic Research • Previous Articles     Next Articles

Isolation of rat Leydig cells with combined low-concentration type Ⅱ and Ⅳ collagenases

Yang Li1, Yu Wang1, Kang Cheng1, Chun Liu1,()   

  1. 1. Department of Urinary Surgery, the First Affiliated Hospital, Shanxi Medical University, Taiyuan 030001, China
  • Received:2017-02-16 Online:2017-07-15 Published:2017-07-15
  • Contact: Chun Liu
  • About author:
    Corresponding author: Liu Chun, Email:

Abstract:

Objective

To establish a simple and effective method to isolate and purify rat Leydig cells.

Methods

Leydig cells were digested with a mixture of 0.1% types II and IV collagenases, purified by differential adherence, and identified by 3β-HSD immunofluorescence staining. The testosterone-secreting capability of Leydig cells was determined by ELISA.

Results

The optimal enzyme digestion time for Leydig cells was about 1 hour. The best adherence time for the purification of Leydig cells was 1-2 h. After 24 hours of culture, the purity of Leydig cells was (98.1 ± 1.5)% as revealed by 3β-HSD immunofluorescence staining, and the survival rate of Leydig cells was (97.3 ± 1.6)%. After incubation for 12 hours, the Leydig cells reached the peak of secretion.

Conclusion

In this study, a simple, stable, and reliable method for isolation and purification of primary Leydig cells was established. This method allows to obtain high-purity Leydig cells with high activity.

Key words: Leydig cell, Isolation, Purification, Identification, Rats

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