Vitrifikasi Blastosis Mencit dengan Metode Kriolupv ?O

• Main Authors: I Wayan Batan, I Ketut Suatha, Wahono Esti PrasetyoningtyaserB, Nining Handayani, Ita Djuwita, Arief Boediono
• Format: Article
• Language: English
• Published: Universitas Udayana 2009-12-01
• Series: Jurnal Veteriner
• Subjects: vitrification, blastocyst, cryoloop
• Online Access: http://ojs.unud.ac.id/index.php/jvet/article/view/3373

<div>Cryopreservation is an ultra rapid freezing process to preserve tissue or organ. The study</div><div>was conducted to identify the effectiveness of cryoloop vitrification method and the viability of</div><div>embryos following vitrification. Embryos at blastocyst stage were vitrified by placing them in</div><div>equilibration medium containing 10% ethylene glycol (EG) and phosphate buffer saline (PBS) wich</div><div>supplemented with 20% new born calf serum for 8-10 minutes. The blastocysts were then removed</div><div>and put in vitrification medium (15% dimethyl sulfoxide, 15% EG, and 0.5M sucrose), and the</div><div>process in the vitrifivcation medium not longer than 25-30 seconds. The blastocysts were immediately</div><div>transferred to the vitrification medium film in the cryoloop and plunged into 100 ml liquid nitrogen.</div><div>The warming process was done by immersing the cryoloop which carried the vitrified blastocsts</div><div>into PBS supplemented with 20% serum and 0.5M sucrose for 1 minute, and then removed to same</div><div>solution supplemented with 0.25M sucrose and 0.1M sucrose for 2 minutes respectively. The</div><div>blastocysts were washed 4 times in kalium simplex optimized medium (KSOM) and cultured in</div><div>drops of KSOM in 5% CO2 incubator at 370C. The observations were done every 6 hours for 48</div><div>hours using inverted microscope ( Olympus IX70 Japan). The viability of embryos was assessed on</div><div>the basis of the intact morphology, reexpansion of the blastosul, and the development of embryos</div><div>into advance stage. The results showed that 85.71% of vitrified embryos, developed into advance</div><div>stages and 19% of them hatched. In conclusion the cryoloop can be used to vitrify the embryos.</div>