Permeability Barrier

Volume 763 of the series Methods in Molecular Biology pp 237-252


An In Vitro System to Study Sertoli Cell Blood-Testis Barrier Dynamics

  • Dolores D. MrukAffiliated withCenter for Biomedical Research, Population Council
  • , C. Yan ChengAffiliated withCenter for Biomedical Research, Population Council Email author 

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The use of an in vitro system based on primary cultures of Sertoli cells isolated from rat testes has greatly facilitated the study of the blood-testis barrier in recent years. Herein, we summarize the detailed procedures on the isolation of undifferentiated Sertoli cells from 20-day-old rat testes, the culture of these cells as a monolayer on Matrigel-coated bicameral units, the characterization of these cultured cells, and the use of the Sertoli cell epithelium for monitoring the integrity of the Sertoli cell blood-testis barrier. This information is based on the routine use of this system in our laboratory to study the Sertoli cell blood-testis barrier in the past two decades, which should be helpful for investigators in the field.

Key words

Testis Sertoli cell Blood-testis barrier Tight junction Ectoplasmic specialization Anchoring junction Spermatogenesis