Efek Penambahan Insulin pada Media Maturasi In Vitro terhadap Pematangan Oosit Sapi Pesisir - Delmita Nugrah Wati; Tinda Afriani; Jaswandi Jaswandi (State Islamic University of Sultan Syarif Kasim Riau, 2023)
ABSTRAK. Penelitian ini bertujuan untuk menentukan kualitas oosit sapi Pesisir yang akan diproduksi secara in vitro dan untuk menentukan dosis penambahan Insulin pada media maturasi in vitro efektif untuk meningkatkan kematangan oosit sapi Pesisir. Sampel yang digunakan yaitu ovarium sapi Pesisir yang diambil di RPH kemudian dilakukan koleksi oosit dengan metode slicing di Laboratorium Bioteknologi. Penelitian ini menggunakan Rancangan Acak Kelompok (RAK) yang terdiri dari 4 perlakuan dan 4 ulangan. Perlakuan yang digunakan adalah penambahan insulin pada media maturasi in vitro yang terdiri dari tanpa insulin (P1), insulin 10 µg/ml (P2), insulin 15 µg/ml (P3) dan insulin 20 µg/ml (P4). Oosit sapi Pesisir dimaturasi dalam media maturasi selama 24 jam di dalam inkubator CO2 dengan suhu 38,5 ᵒC. Sampel yang sudah dimaturasi diwarnai dengan 2% aceto-orcein selama 5 menit kemudian zat pewarna dihilangkan dengan campuran (asam asetat, gliserin dan aquadest) selanjutnya pengamatan inti diamati dengan menggunakan mikroskop. Hasil penelitian menunjukkan bahwa oosit yang paling banyak ditemukan adalah oosit grade B (75,59%), A(13,53%), C (8,48%), dan yang paling sedikit adalah grade D (2,40%). Penambahan Insulin dengan dosis 10 µg/ml dan 15 µg/ml pada media maturasi in vitro dapat meningkatkan pematangan oosit sapi pesisir yang mencapai metaphase II (MII) daripada tanpa insulin, sedangkan penambahan dosis insulin 20 µg/ml menurunkan pematangan oosit sapi pesisir yang mencapai MII. Berdasarkan hasil penelitian tersebut disimpulkan bahwa oosit yang dimaturasi dengan penambahan insulin 10 µg/ml dan 15 µg/ml sangat berpengaruh (P<0,01) terhadap pematangan oosit mencapai MII dan hasil terbaik adalah penambahan insulin 15 µg/ml sebesar 84,58%. Kata kunci: Insulin, Oosit Sapi Pesisir, Pematangan Inti.Effect of Insulin Addition on In Vitro Maturation Media on Pesisir Cattle Oocyte MaturationABSTRACT. This study aims to determine the quality of Pesisir cattle oocytes to be produced in vitro and to determine that adding different insulin to in vitro maturation media effectively increases the maturity of pesisir cattle oocytes. The sample was bovine ovaries of Pesisir cattle taken at the RPH and then bovine oocytes collected at the Biotechnology Laboratory. This study used a randomized block design (RBD) which consisted of 4 treatments and 4 replications. The treatment used was adding of insulin to in vitro maturation media consisting of no insulin (P1), insulin 10 µg/ml (P2), insulin 15 µg/ml (P3) and insulin 20 µg/ml (P4). Pesisir bovine oocytes were matured in maturation medium for 24 hours in a CO2 incubator with a temperature of 38.5 ᵒC. Samples that had been matured were stained with 2% aceto-orcein for 5 minutes and then the dye was removed with a mixture (acetic acid, glycerin and distilled water) then the observations of the nucleus were observed using a microscope. The results showed that the most common oocytes were grade B oocytes (75.59%), A (13.53%), C (8.48%), and the fewest were grade D (2.40%). The addition of insulin at a dose of 10 µg/ml and 15 µg/ml in in vitro maturation media could increase the oocyte maturation of coastal bovines that reached metaphase II (MII) than without insulin, while the addition of 20 µg/ml insulin decreased the oocyte maturation of coastal bovines that reached MII. Based on the results of this study it was concluded that oocytes which were matured with the addition of insulin 10 µg/ml and 15 µg/ml had a very significant effect (P<0.01) on oocyte maturation reaching MII and the best treatmentin this study was an insulin dose of 15 µg/ml of 84.58%.