Isolation of Immune Regulatory Sertoli Cells from Mouse Seminiferous Tubules

Published: August 31, 2023

Abstract

Source: Bhushan, S. et al., Isolation of Sertoli Cells and Peritubular Cells from Rat Testes. J. Vis. Exp. (2016).

This video demonstrates the isolation procedure of Sertoli cells from mouse seminiferous tubules, a component of the testes. Enzymatic digestion and repeated washes promote the release of Sertoli cells into the solution, which can be cultured further for proliferation and growth.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Preparation of Media, Enzyme Solutions, and Animals Prepare 1% iodine alcohol by dissolving 1 g of iodine in 100 ml ethanol. Prepare 2x 500 ml Dulbecco´s phosphate buffered saline (PBS) without Ca2+ and Mg2+ supplemented each with 7.5 ml D-glucose (100 g/L) and 5 ml 100x Pe…

Representative Results

Figure 1. Testes from Wistar rats are excised and decapsulated. (A) The peritoneal cavity is opened through a longitudinal incision as described in the text. An asterisk (*) indicates the epidydimal fat pad. (B) Testes are removed by cutting the spermatic cord and (C) collected in a 50 ml conical tube containing 20 ml PBS. When all testes have been collected they are d…

開示

The authors have nothing to disclose.

Materials

Corning™ Cell Strainers 70 µm Corning 431751 White color
DAPI mountant ProLong® Gold Antifade Life Technologies P-36931
D-glucose Sigma G8644 100 g/l
Dulbecco's PBS without Ca2+/Mg2+ Gibco 14190-094
DNase I Roche 10104159001
Hyaluronidase from bovine testis Sigma H3506
Penicillin (5,000 U/ml) Streptomycin (5,000 µg/ml) Gibco 15070-063 100x solution
RPMI-1640 Gibco 21875-034 Contains 300 mg/L L-glutamine
Trypsin from porcine pancreas Sigma T5266
Trypan Blue Stain (0.4%) Gibco 15250-061
Trypsin inhibitor from soybean Sigma T6522 The Sigma product is considerably cheaper than the previously used BPTI (Aprotinin) from Roche.
Wistar WU rats Charles River N/A Should be 19 days old on day of experiment.

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記事を引用
Isolation of Immune Regulatory Sertoli Cells from Mouse Seminiferous Tubules. J. Vis. Exp. (Pending Publication), e21562, doi: (2023).

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