TRANSPLANTATION OF SPERMATOGONIA ISOLATED FROM GIANT GOURAMI COLD PRESERVED TESTIS INTO NILE TILAPIA LARVAE

The recent study has been conducted to develop testicular germ cell (TGC) transplantation as a tool for preservation and propagation of male germ-plasm from endangered fish species. In practice of TGC transplantation, recipient and donor cell may not be immediately available at the same time whereas the testis can not be survive longer when it is outside of the body. Therefore, preservation of testis tissue may be required before transplantation. The research was conducted to evaluate 1) the viability of spermatogonia isolated from short term preserved testisand 2) colonization efficiency of preserved donor cells after transplantation.Testis was preserved in physiological NaCl solution at 4 o C for 6, 12,24 and48 hours. Testis were dissociated in 0.5 % trypsin and 3% DNase 10 IU/μL in PBS(phosphate buffered solution) complemented with 5% FBS ( fetal bovine serum), 25 mM HEPESand 1mM CaCl2 to obtain testicular germ cell suspension. Testicular germ cell isolated from 24 and 48 hours preservation and labeled with PKH 26 membrane fluorescent dye then transplanted into 3 days post hatched tilapia recipient. The results showed that the viability of spermatogonia started to decrease significantly in 12 hours preservation (P<0.05)and in 48 hours preservation, the amount of viable cells was only 54,48±8,33%. Two months post transplantation, the efficiency of colonizationwere analyzed in recipient and the result showed insignificant difference of efficiency colonization between recipient transplanted with testicular tissue preserved 24 and 48 hours (55,56% each) and without preservation (61,11%) . In conclusion, preserved testicular tissue at 4 o C could be used as the source of donor cell for testicular germ cell transplantation of giant gourami into Nile tilapia.