MORPHOLOGICAL CHARACTERISTIC OF SPERMATOGONIA AND TESTES DISSOCIATION : A Preliminary Study for the Germ Cell Transplantation in Giant Gouramy (Osphronemus gouramy)
Abstract: The recent study
were attempting to develop spermatogonial germ cell transplantation as a tool
to preserve and propagate male germ-plasm from endangered fish species, as well
as to produce surrogate broodstock of commercially valuable fish. Spermatogonia
identification and testes dissociation were the first necessary steps to obtain
highly amount and viable population of spermatogonia as donor cells for
transplantation. Using giant gouramy testes as a model, spermatogonia was
histological characterized and two methods of testes dissociations were
compared (i.e. medium A contained 0.5% trypsin in PBS and medium B contained
0.5% trypsin and DNase 10 IU/μL in PBS complemented with CaCl2, Hepes and FCS).
Optimal incubation times (1, 2, 3, 4 and 5 hours) in dissociation medium were
also determined. Freshly isolated testes of immature giant gouramy were minced
in dissociation medium and then incubated to get monodisperce cell suspension.
Parameters observed were number and viability of spermatogonia (ø > 10 μm).
The viability was analyzed using trypan blue exclusion dye. The results showed
that the average number of spermatogonia observed in medium B was higher than
in medium A (P<0.05), meanwhile the viability of spermatogonia between
medium A and B were not significantly different (P>0.05). The viability of
spermatogonia decreased by the increasing duration time of dissociation. The
viability of spermatogonia started to decrease significantly in 2 hours
incubation time in medium A and 4 hours incubation time in medium B
(P<0.05). In conclusion, application of dissociation medium B yielded higher
number of viable spermatogonia than dissociation medium A.
Keywords: testes;
spermatogonia; morphology; dissociation; viability; giant gouramy
Author: Irma Andriani, Ita
Djuwita, Komar Sumantadinta, Muhammad Zairin Junior, Harton Arfah, Alimuddin
Journal Code: pperikanangg100026
