Isolation of Immune Regulatory Sertoli Cells from Mouse Seminiferous Tubules

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Samenvatting

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 Penicillin/Streptomycin stock solution (15 mg/ml D-glucose, 50 U/ml Penicillin and 50 µg/ml Streptomycin final).
Supplement 1x 500 ml Ros....

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Resultaten

Rodent ovary dissection, tissue isolation, and preservation for further analysis, experiment steps shown.
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 sperm.......