PENGARUH PENAMBAHAN Insulin-Transferin Selenium (ITS) PADA MEDIA KULTUR PEMATANGAN OOSIT SAPI POTONG SECARA IN VITRO

MIA SARI HIDAYATI, drh. Yuda Heru Fibrianto MP., P.hD

2015 | Skripsi | S1 KEDOKTERAN HEWAN

Penelitian ini bertujuan untuk mengetahui pengaruh Insulin-Transferin Selenium (ITS) pada pematangan oosit sapi potong secara in vitro. Oosit sapi potong dikoleksi dari Rumah Potong Hewan (RPH) Kota Yogyakarta dan diletakkan dalam termos berisi air hangat 37o C sebanyak 454 ml dengan diberi 0,88% NaCl dan 0,11% Amoxicillin lalu dilakukan perlakuan di Laboratorium IVF, Rumah Sakit Gladiool Magelang. Isi folikel pada oosit termaksud cumulus-oocytes complexes (COC's), yang dikoleksi dengan aspirasi pada antrum follikel kecil yang terlihat (diameter sekitar 2-6 mm) menggunakan jarum suntik 10 ml dengan needle berukuran 18 Gauge. Oosit dikultur pada dua media , yang pertama media kultur yang berisi 1000 µl DMEM , 10% Serum FBS sebanyak 10 µl, 5% FSH sebanyak 5 µl, pennicilin 600 IU/ml , Streptomycin 100 mg/ml selaku kontrol dan untuk media kultur sebagai perlakuan ditambahkan dari bahan tadi 1% ITS sebanyak 1 µl pada media culture selama minimal 24 jam pada suhu 37o C di inkubator 5 % Co2. Pengamatan dilakukan dengan mikroskop stereo dan mikromanipulator system dilakukan pengamatan polar body dan ekspansi cumulus pada oosit pasca inkubasi. Hasil penelitian ini diuji secara statistika menggunakan 2, menunjukkan adanya asosiasi penambahan Insulin-Transferrin Selenium (ITS) pada media kultur pematangan oosit sapi potong secara In Vitro Maturation terhadap tingkat kematangannya ( 2= 5,523; P= 0,019; OR= 0,475). Tingkat kematangan Oosit sapi potong pada media kultur dengan penambahan ITS 2,1 kali lebih tinggi tingkat kematanganya. Pengulangan sebanyak 4 kali menunjukkan pada media kultur menggunakan ITS presentase kematangannya 88,2 %; 50 %; 100 %; 51,85% dan media kultur tanpa ITS 86,67 %; 20 %; 72,73 %; 39,74 %. Kesimpulannya, pemberian Insulin-Transferrin Selenium (ITS) memiliki pengaruh terhadap peningkatan angka kematangan oosit pada media kultur secara In Vitro.

The aim of the present study was to evaluate the effect of Insulin-Transferin Selenium (ITS) in media culture of cattle oocyte on maturation in vitro. Cattle ovary were collected at a local slaughterhouse in Yogyakarta and put in thermos bottle contain warm water with 37o C of temperature and be given inside it 4 gr of NaCl ; 500 mg Amoxillin and the treatment of this research we do in IVF laboratory, Gladiool Hospital of Magelang. The follicular content including cumulus-oocyte complexes (COC's), were colected by aspirating the visible small antral follicles (around 2-6 mm diameter) with 10 ml syringe equipped with an 18 gauge needle. Oocytes were cultured in two medium, first culture medium is contain 1000 µl DMEM, 10% FBS 10 µl, 5% FSH 5 µl, pennicilin 600 IU/ml, Streptomycin 100 mg/ml as control and for the second culture medium as Treatment same as control medium but adding 1% of ITS 1 µl then incubation minimum for 24-hours at 37o C on 5% CO2 inkubator. The observation with microskop stereo equipment and system manipulator equipment for observe the polar body and expanded of cumulus in oocyte after inkubated. The result in statistics test that using x2 showed there is association in adding Insulin-Transferrin Selenium (ITS) maturation culture media in vitro toward of oocyte mature level (x2= 5,523; P= 0,019; OR= 0,475). Level of cattle oocyte mature media with ITS 2,1 more high in mature level. This research that do with 4 replication showed in culture medium that using ITS have maturation percentage are 88,2 %; 50 %; 100 %; 51,85% and for culture medium without ITS the maturation percentage are 86,67 %; 20 %; 72,73 %; 39,74 %. The conclusion, adding Insulin-Transferrin Selenium (ITS) in culture media of oocyte in IVM could effect increase a number mature of oocyte.

Kata Kunci : Oocyte, Cattle, In Vitro Maturation, Insulin Transferin Selenium, Media