Perbandingan Viabilitas Oosit Domba Pasca Vitrifikasi dengan Menggunakan Hemistraw dan Cryotop
Abstract
ABSTRAK. Tujuan dari penelitian ini adalah untuk mengkaji efek vitrifikasi dengan menggunakan dua buah system carrier yang berbeda terhadap viabilitas oosit domba yang telah dimaturasi secara in vitro. Oosit dibagi menjadi dua kelompok perlakuan, yaitu (i) divitrifikasi dengan menggunakan hemistraw (ii) divitrifikasi dengan menggunakan cryotop. Viabilitas oosit dievaluasi berdasarkan reekspansi, warna dan homogenitas sitoplasma. Hasil penelitian ini menunjukkan bahwa viabilitas oosit setelah vitrifikasi serupa pada kedua jenis carrier yang digunakan untuk vitrifikasi. Oosit diletakkan dalam larutan equilibrasi yang mengandung konsentrasi permeable krioprotektan setengah dari larutan vitrifikasi. Setelah 15 menit, oosit ditransfer ke dalam media vitrifikasi yang mengandung 17% EG+17% DMSO +0, 65M sukrosa di dalam modified PBS yang disuplementasi dengan 20% fetal bovine serum. Total waktu yang digunakan untuk memaparkan oosit ke dalam larutan vitrifikasi adalah 30 detik. 5-8 oosit dipipet menggunakan kapiler gelas dan diletakkan/loading ke dalam carrier yang digunakan (hemistraw atau cryotop) kemudian langsung di paparkan ke dalam nitrogen cair. Viabilitas oosit dievaluasi berdasarkan reekspansi, warna dan homogenitas sitoplasma. Hasil penelitian ini menunjukkan bahwa viabilitas oosit setelah vitrifikasi serupa pada kedua jenis carrier yang digunakan untuk vitrifikasi
(Comparation of sheep oocyte viability after vitrification using hemistraw and cryotop)
ABSTRACT. The aim of this study was to examine the effect vitrification using two different carrier system on the matured sheep oocytes viability. Oocytes were devided into two group (i) vitrified using hemistraw (ii) vitrified using cryotop. Oocytes placed into equilibration solution which is containing a half concentration permeable cryoprotectant of vitrification solution. After 15 minute, oocytes were transferred into vitrification solution containing 17% EG+17% DMSO +0, 65M sucrose in modified PBS supplemented with 20% fetal bovine serum. The total exposure time of oocytes to vitrification solution was 30 sec. Oocytes were pipetted into a glass capillary into group 5-8, and loaded into carrier (hemistraw or cryotop) then plugged into liquid nitrogen. After a week cryopreservation, oocytes were warmed and cultured in TCM 199 suplemented with 10% fetal bovine serum at 38.50 C under 5% CO2 for 3h. Oocytes viability was evaluated by re expansion, color and homogeneity of oocyte cytoplasm. Our results indicated that the oocytes viability after vitrification was similar from both of carrier for vitrification
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DOI: https://doi.org/10.17969/agripet.v17i2.8131
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