Human Reproduction, Vol. 14, No. 5, 1287-1293,
May 1999
© 1999 European Society of Human Reproduction and Embryology
In-vitro maturation of round spermatids using co-culture on Vero cells
1 Instituto Bernabeu de Fertilidad y Ginecologia, Alicante, Spain, 2 Department of Medical Genetics, Faculty of Medicine and 3 Laboratory of Cell Biology, Institute of Biomedical Sciences, University of Porto, Lg. Prof. Abel Salazar 2, 4050 Portugal
In an attempt to determine whether co-culture could promote sperm maturation, three patients with non-obstructive azoospermia, two with maturation arrest at the level of primary spermatocytes and one patient with <1% tubules showing complete spermatogenesis, and one patient with total globozoospermia, gave consent to experimentally co-culture round spermatids retrieved from the testicle on Vero cell monolayers. In all azoospermic patients elongating spermatids could be obtained from round spermatids. In one case of maturation arrest, of 37 round spermatids co-cultured for up to 5 days, 30% developed flagella, 46% matured to elongating and 19% to elongated spermatids, with one mature spermatozoon also obtained (3%). In the same patient, primary cultures of three round spermatids with flagella enabled development of one further mature spermatozoon. In the case with total globozoospermia, of six round spermatids co-cultured for up to 5 days, one mature spermatozoon was obtained, with a flagellum and normal head morphology. These preliminary findings suggest that it may be possible to overcome the round spermatid block, and even the triggering of morphological abnormalities arising at the spermiogenic level, by in-vitro maturation under special environmental conditions.
Key words: co-culture/in-vitro germ cell maturation/non-obstructive azoospermia/round spermatids/Vero cells
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