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Rat sertoli cells: a rapid method for obtaining viable cells.

M J Welsh, J P Wiebe

|March 1, 1975

Summary

This study details a rapid 3-hour method for isolating viable Sertoli cells from rat testes using enzymatic digestion and density gradient sedimentation. The enriched cell populations achieve high purity and viability, suitable for further research.

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Area of Science:

Reproductive Biology
Cell Biology
Animal Science

Background:

Sertoli cells are crucial for spermatogenesis and male fertility.
Efficient isolation of pure, viable Sertoli cells is essential for research.
Current methods may be time-consuming or yield lower purity.

Purpose of the Study:

To develop a rapid and efficient method for isolating viable Sertoli cells from rat testes.
To achieve high purity and enrichment of Sertoli cells.
To provide a reliable cell population for in vitro studies.

Main Methods:

Sequential enzymatic digestion of minced rat testes (15-29 days old) with collagenase and pancreatin.
Separation of cells using sucrose density gradient sedimentation.
Assessment of cell viability using trypan blue exclusion and in vitro culturing.

Main Results:

Obtained Sertoli cell populations with 60-82% purity, representing a 2-5 fold enrichment.
Preparations were free of interstitial cells and largely free of peritubular cells.
Sertoli cells demonstrated 95% viability, with high purity (95-98%) achieved in monolayer cultures within 3 days.

Conclusions:

The described method provides a fast (3-hour) and effective way to isolate highly viable and enriched Sertoli cell populations.
This technique is suitable for generating high-purity Sertoli cell cultures for reproductive biology research.
The method offers a significant improvement over existing protocols for Sertoli cell isolation.

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