Leptin supplementation in embryo culture medium increases in vivo implantation rates in mice

Abstract

All mice experiments and animal care protocols were approved by Koç University Local Ethics Committee for Animal Experiments (approval number: 2013-06). The animals were kept in Koç University, Animal Research Facility of Center for Translational Medicine (KUTTAM) under 12 h light-12 h dark cycle, and a diet of commercial pellet food ad libitum and automatic water containers were provided. 2.1. Embryo collection CB6F1 (C57BL⁄6j ͯ BALB⁄c) female mice (n = 24) were intraperitoneally injected (IP) with 10 IU of equine Abstract: Leptin is a hormone-like protein consisting of 167 amino acids. The aim of this study is to compare the effects of different leptin concentrations on in vitro and in vivo embryo development rates. In vitro development rates were investigated by embryo culture studies, and in vivo implantation rates and the quality of embryos were assessed by embryo transfers to the recipient mice. The results showed that addition of leptin into the embryo culture medium at 10 and 100 ng/mL doses supported the in vitro development of mouse embryo. Moreover, leptin increased the total cell number of blastocyst, particularly the trophectoderm cells. In vivo assessment showed a significant increase in the proportion of the embryos implanted in 10 and 100 ng/mL groups, compared to the control group. In conclusion, leptin supplement in embryo culture medium increases implantation rates in mice.

Comparison of parthenogenetic oocyte activation in different mouse strains on in vitro development rate and quality

“…The oocytes were derived from rupturing oviduct ampulla and washed in Human Tubal Fluid + HEPES buffered (HTF, global total w / HEPES) medium + 80 IU/mL hyaluronidase (Sigma H -3506) + 4 mg/mL Bovine Serum Albumin (BSA, Fraction V. Sigma A3311) and isolated oocyte washed three times in 500 µl HTF medium and selected only high-quality mouse oocytes. Then, oocytes were transferred into four well plates (18)(19)(20)(21).…”

“…At least 2 hours prior, embryo culture media were incubated at 5% CO2 and 37°C temperature and high humidified in an incubator for gassing. Oocytes were cultured at embryo culture medium (4 mg/mL BSA) for 120 hours until blastocyst stage (18)(19)(20)(21).…”

Effects of different parthenogenetic activation periods on mouse embryo development and quality

“…On the following day, the blastocysts were transferred to the glycerol droplet after washing in 5 μl glycerol droplet on each glass slide, and covered with a coverslip for blastocyst stabilization. The blastocyst preparations were observed using an inverted microscope with a red and blue fluorescence attachment for the determination of trophectoderm (TE) and inner cell mass (ICM) cell numbers (Mallol et al 2013, Taşkın et al 2019b.…”