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1 Institute of Histology and
General Embryology, University of Rome,
Rome, Italy
A new technique involving a brief hypotonic
treatment was developed for obtaining pure rat
Sertoli cell cultures. This method for the selective removal of the germ cells present in Sertoli
cell enriched cultures (SCEC) is based on the
differential response of the two cell types to
changes in osmolarity. It was found that the
optimal conditions for germ cell detachment
without Sertoli cell impairment consist of incubation for 2.5 minutes at 20 C in 20 mM TRIS-HCl. When compared with SCEC, the Sertolicell-only cultures (SCOC) thus obtained retain
unaltered morphologic features and responsiveness to FSH stimulation (morphologic modifications and 17 
-estradiol secretion). The
availability of pure Sertoli cell cultures (ie, free
of contaminating germ cells) provides an advantage in the study of their metabolic activity.
Moreover, with this technique it is feasible to
compare Sertoli cell function in association
with germ cells to function in the absence of
germ cells.
Key words: cell cultures, Sertoli cell, germ cell
Submitted on October 6, 1980
Revised on January 28, 1981
Accepted on February 25, 1981
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