PRODUKSI EMBRIO KUCING SECARA IN VITRO DARI SPERMATOZOA HASIL PRESERVASI MELALUI FERTILISASI MIKRO
(In Vitro Development of Cat Embryos from Preserved Sperm by Micro Fertilization)
Kartini Eriani, Yuhara Sukra, Arief Boediono, Ita Djuwita, Sony Heru Sumarsono
ABSTRAK
Penelitian ini bertujuan mengetahui motilitas dan kemampuan memfertilisasi sperma kucing yang berasal dari epididimis yang disimpan pada suhu 4 °C. Epididimis disimpan dalam media phosphate buffer saline (PBS) pada suhu 4 C selama 1, 3, dan 6 hari. Viabilitas spermatozoa diamati dengan pewarnaan hoechst-propidium iodine. Fungsi biologis spermatozoa dievaluasi melalui teknik kultur in vitro dengan fertilisasi mikro dan perkembangan embrio di dalam media kultur CR1aa. Hasil penelitian menunjukkan persentase spermatozoa hidup pada hari ke-1, 3, dan 6 penyimpanan masing-masing adalah 81,0; 71,7; dan 70,7% (duktus deferens), 84,0; 81,2; dan 63,2% (kauda epididimis), 84,0%; 75,0; dan 74,7% (korpus epididimis). Persentase pronukleus (PN) yang didapat dengan teknik intra cytoplasmic sperm injection (ICSI) menggunakan spermatozoa epididimis pada hari ke-1, 3, dan 6 hari penyimpanan masing-masing adalah 8,0; 10,0; dan 5,9%. Preservasi epididimis pada suhu 4 oC dalam PBS dapat digunakan untuk fertilisasi dan menghasilkan embrio kucing secara in vitro.
Kata kunci: gamet, preservation (penyimpanan), fertilisasi in vitro, intracytoplasmic sperm injection (ICSI)
ABSTRACT
The purpose of this study was to determine how long cat’s spermatozoa remained motile when maintained in epidydimis stored at 4 °C. Epidyidimis was preserved immediately in phosphate buffer saline at 4 oC for 1, 3, and 6 days. The observation on sperm viability after preservation were identified through Hoechst-Propidium Iodine. The sperm biological function was evaluated using in vitro culture technique for micro fertilization and embryonic development rate in CR1aa medium culture. The results of the study showed that the live sperm percentage of sperm collected from ductus deferens, cauda, and corpus epidydimis from 1, 3, and 6 days of preservation times were 81.0, 71.7, and 70.%7; 84.0, 81.2, and 63.2%; 84.0, 75.0, and 74.7%, respectively. The percentage of pronucleus produced from ICSI using epidydimal sperm after preserved 1, 3,and 6 days resulted in 8.0, 10.0, and 5.9%, respectively. In conclusion, preservation of epididyimis at 4 oC in PBS can be used to IVF and in vitro production of cat embryos.
Key words: gamet, preservation, in vitro fertilization, intracytoplasmic sperm injection (ICSI)
Publish in Jurnal Kedokteran Hewan Vol. 7 No. 1, Maret 2013 p 37-42