Human Reproduction, Vol 12, 1682-1687, Copyright © 1997 by Oxford University Press
E Crabbe, G Verheyen, H Tournaye and A Van Steirteghem
Intracytoplasmic sperm injection (ICSI) with testicular spermatozoa and
ejaculated spermatids has been applied successfully to treat infertility
because of azoospermia due to obstruction or testicular failure. In most
cases, free spermatozoa are recovered from testicular tissue after
mechanical mincing of multiple biopsies. Testicular sperm retrieval,
however, remains unsuccessful in a high proportion of male patients
suffering from testicular failure. In the present study, four media for
enzymatic collagen digestion of testicular tissue were compared on 15
testis biopsies obtained from azoospermic patients following an ICSI
treatment and on five biopsies from orchiectomy patients. To dissociate the
testicular tissue, collagenase type IA, collagenase type IV, collagenase
type IA supplemented with elastase and collagenase type IV supplemented
with elastase were used. The preparations with only collagenase type IV
provided the best dissolution of the cells from their tissue, with a higher
yield of free vital testicular spermatozoa and single round germ cells.
Enzymatic digestion of testicular tissue has the advantage, especially in
cases of testicular failure, that several biopsies can be concentrated into
one pellet of single cells. In this way, the sperm retrieval in cases of
testicular failure might be optimized to yield the maximal numbers of
spermatids and/or spermatozoa.
ARTICLES
The use of enzymatic procedures to recover testicular germ cells
Centre for Reproductive Medicine, University Hospital, Dutch-speaking Brussels Free University, Belgium.
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