VIABILITAS OOSIT DOMBA PASCATRANSPLANTASI OVARIUM DOMBA DALAM UTERUS KELINCI PSEUDOPREGNANT

Abstract

ABSTRAK Telah dilakukan penelitian untuk mengetahui viabilitas oosit domba yang dikoleksi dari ovarium domba pascatransplantasi intrauterus pada kelinci pseudopregnant. Ovarium domba ditransplantasikan dalam uterus kelinci pseudopregnant pada hari pertama bunting semu dan kemudian diambil kembali setelah lima (P5) atau tujuh (P7) hari transplantasi. Sebagai kontrol digunakan ovarium segar. Oosit dikoleksi dari ovarium pascatransplantasi dengan metode slicing (pencacahan) di dalam media phosphate buffer saline (PBS) yang disuplementasi dengan 5% fetal bovine serum (FBS) dan 100 IU penicillin-streptomisin/ml. Oosit hasil koleksi selanjutnya dimaturasi dalam Tissue Culture Medium (TCM)-199 yang disuplementasi dengan 10% FBS, 10 IU follicle stimulating hormone (FSH)/ml dan 100 IU penicillin-streptomisin/ml. Oosit selanjutnya diinkubasi dalam inkubator CO2, dengan kandungan CO2 5% dan suhu 38ºC, selama 24 jam. Setelah dimaturasi, oosit diwarnai dengan aceto-orcein 2% untuk menentukan status inti oosit. Hasilnya memperlihatkan bahwa oosit yang mampu mencapai tahapan perkembangan dengan status inti M-II setelah maturasi pada P5 (35,05%) dan P7 (35,24%) nyata lebih sedikit (p<0,05) dibandingkan dengan kontrol (56,65%). Dapat disimpulkan bahwa viabilitas oosit domba pascatransplantasi di dalam uterus kelinci pseudopregnant masih ditemukan meskipun persentasenya lebih rendah. EWE OOCYTE VIABILITY FROM EWE OVARIAN AFTER INTRAUTERINE TRANSPLANTATION TO PSEUDOPREGNANT RABBIT ABSTRACT The aim of the present study was to investigate the ewe oocyte viability from ewe ovary after intrauterine transplantation to pseudopregnant rabbit. The ewe ovary was transplanted intrauterine on day 1 to pseudopregnant rabbit and oocytes recollected on five (P5) or seven (P7) days posttransplantation. The fresh ovary was used as the control. The oocytes were collected from the ovaries by slicing method in Phosphate Buffer Saline (PBS) supplemented with 5% of Fetal Bovine Serum (FBS), and 100 IU/ml of penicillin-streptomycin. Oocytes were matured in Tissue Culture Medium (TCM)-199 supplemented with 10% FBS, 10 IU/ml of Follicle Stimulating Hormone (FSH), and 100 IU/ml of penicillin-streptomycin. Oocytes were incubated in CO2 incubator with 5% CO2, 38°C for 24 h. After maturation, the oocytes were stained with 2% aceto-orcein to determine the nuclear oocytes status. The result showed that the oocytes could reach the M-II phase from P5 (35.05%) and P7 (35.24%) decreased significantly (p<0.05) compared to the control (56.65%). However it can be concluded that the oocytes viability still preserved intrauterine in pseudopregnant rabbit.