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Endocrinology, doi:10.1210/endo-109-2-667
Endocrinology Vol. 109, No. 2 667-669
Copyright © 1981 by the Endocrine Society.
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ISOLATION OF PURIFIED RAT LEYDIG CELLS USING CONTINUOUS PERCOLL GRADIENTS

JOAN Y. BROWNING, ROSARIO D'AGATA1 and H. EDWARD GROTJAN, JR.

Department of Reproductive Medicine and Biology, University of Texas Medical School, P. 0. Box 20708, Houston, Texas 77025

Abstract

Continuous, isoosmotic Percoll gradients were utilized to isolate highly purified (90–95%) Leydig cells from rat testicular interstitial cell suspensions. The purified Leydig cells were contained in the third of four bands generated on the gradient, and retained their morphologial and biochemical integrity. The cells in the purified Leydig cell fraction stained positively for 3β-hydroxysteroid dehydrogenase activity, appeared characteristic of steroid synthesizing cells when examined electron microscopically, specifically bound 125I_hCG and responded to hCG stimulation in vitro by producing testosterone. This procedure for isolating purified Leydig cells has advantages over methods currently in use as the gradients are essentially isoosmotic throughout, are easily generated, relatively inexpensive to use, and are readily applicable to sterile procedures when cell culture is anticipated.

Footnotes

1 Present address: NATO Senior Fellow (CNR), Department of Medical Pathology, University of Catania, Catania, Italy.




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Copyright © 1981 by The Endocrine Society