Histological description of aceh cattle ovary cryopreserved by various cryoprotectants

Abstract

This research aimed to determine Aceh cattle ovarian follicle morphological integrity after vitrified by various cryoprotectants. Cryoprotectants used in this research were 30% ethylene glycol (EG), 30% dimethyl suphocide (DMSO), and combination of 15% EG + 15% DMSO. Prior to vitrification process, ovaries were cleansed by phosphate buffered saline (PBS) and then cut into ±1 mm³. Ovaries were consecutively submerged into the following liquid for 5 minutes each: PBS+ 0.25 M sucrose; PBS+ 0.5 M sucrose; PBS+ 0.5 M sucrose + 10% cryoprotectants; and PBS+ 0.5 M sucrose + 30% cryoprotectants. Then, ovaries were packed into straws with ±7 cm in length and ± 6 mm in diameter. Before kept in liquid nitrogen, ovaries were first exposed to nitrogen fume for 10 second. After being stored for 1 day, the ovaries were proceed for histological examination. The result showed that Aceh cattle ovarian follicle after vitrification using 30% EG yields the best morphological integrity. Cumulus oophorus, zona pellucida, granulose cell arrangement, theca interna, and theca externa cells were observed clearer in ovary which was vitrified with 30 % EG than those with 30% DMSO and combination of 15% EG + 15% DMSO. As conclusion, 30% EG was able to protect ovary morphological integrity better than 15 % EG + 15% DMSO and 30% DMSO. Furthermore, combination of 15% EG+ 15 % DMSO was relatively better in protecting ovary follicle morphological integrity compared to 30% DMSO.