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BOR - Papers in Press, published online ahead of print November 27, 2002.
Biol Reprod 2002, 10.1095/biolreprod.102.008045
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Submitted June 4, 2002
Returned for revision July 1, 2002
Accepted October 3, 2002

Testis


Isolation of Sertoli Cells from Adult Rat Testes: An Approach to Ex Vivo Studies of Sertoli Cell Function

Matthew D. Anway 1*, Janet Folmer 1, William W. Wright 1, Barry R. Zirkin 1
1 Johns Hopkins University

* To whom correspondence should be addressed. E-mail: manway{at}jhsph.edu.

Abstract

Much of what is known about the molecular regulation and function of adult Sertoli cells has been inferred from in vitro studies of immature Sertoli cells. However, adult and immature cells differ in significant ways and, moreover, many Sertoli cell functions are regulated by conditions that are difficult to replicate in vitro. Our objective was to develop a procedure to isolate Sertoli cells rapidly and purely enough to make it possible to treat rats in vivo and then assess Sertoli cell function immediately after the isolation of the cells. The isolation procedure described herein takes approximately 4 hours. From a single testis of a young adult rat (age 4 months), we obtain 7.0±0.4 x106 Sertoli cells; and from old rats (age 21 months), 7.2±0.4 x106 Sertoli cells. The purity, determined by morphological analyses of plastic- embedded cells or after staining for tyrosine-tubulin or vimentin, averaged about 80%. The contaminants typically included germ cells (10%)and myoid cells (10%). The germ cell-expressed genes protamine-2 and hemiferrin were not detected in the Sertoli cell preparations by Northern blot analyses; the Sertoli cell-expressed genes clusterin and transferrin were highly expressed.



Key words: Aging • Sertoli cells • Spermatogenesis


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