Kualitas, Kemampuan Implantasi dan Viabilitas in-vivo Embrio Mencit (Mus musculus) Galur Swiss Webster setelah Pembekuan dengan Metode Vitrifikasi

Abstract

Reproductive technology including in vitro fertilization (IVF), embryo manipulation, gamete and embryo freezing, thawing and embryo transfer were rapidly developed. Vitrification is an embryo freezing technique that most developed. In this experiment, we vitrified mouse embryos and then examined the embryos i.e: (i) the quality of the embryos after thawing, (ii) the implantation rate of the embryos and (iii) viability of the embryos in vivo. Morula and blastocycst were collected from female mice that were pregnant day 3.5. The embryos were equilibraten in mPBS +10% ethylene glycol. Vitrification was carried out by a drop of VABEDS medium containing 6-10 embryos into a tip of a straw then frozen in liquid nitrogen for 24 hours.Thawing was carried out by flushing the ernbryos by using mPBS suplemented with 0.5, 0.25, 0.1 and 0 M sucrose. After incubated in M2 medium at 37 0c for 1-2 hours, the recovery embryos were then transferred into the uteri of day 2.5 of pseudopregnant female. The female were then sacrificed at day 16 of gestation and the total implantation, total life and death fetuses, and resorpted embryos were taken as data. The results showed that vitrification significantly (p<0,05) reduce the quality of the embryos, as well as their implantation rate and the viability of the fetuses which may be caused by the unoptimal combination of the cryoprotectant in the vitrification medium, temperature and exposure time during vitrification