In vitro and in vivo quality of bovine embryos in vitro produced with sex-sorted sperm

Abstract

In this work we analyzed the effects of three culture systems on developmental ability of bovine embryos in vitro produced with sexed sperm, the survival to vitrification (cryologic vitrification method) of such blastocysts, and their pregnancy rates after embryo transfer to recipients, both as fresh and after vitrification/warming. Finally, we measured the accuracy of the sorting protocol by a polymerase chain reaction-based method to validate the embryo sex at blastocyst stages. We confirmed an individual effect of the bull as well as development rates of embryos produced with sorted sperm lower than embryos with unsorted sperm, independent of the culture system used. The cryoresistance to vitrification of embryos produced with sexed sperm did not differ from that of conventionally produced embryos (re-expansion rates at 24 and 48 h: 74.6% vs. 75.5%, and 64.5% vs. 68.1% for embryos produced with conventional and sorted sperm, respectively; hatching rates at 48 h: 63.55% vs. 55.5% for embryos produced with conventional and sorted sperm, respectively). Finally, no significant differences were found in pregnancy rates after the embryo transfer of fresh and vitrified/warmed blastocysts (52.8% vs. 42.0%, respectively; P Ͼ 0.05). Male and female embryos produced with sorted sperm showed the same quality in terms of developmental ability, cryoresistance, and pregnancy rates after transfer. Our culture system, coupled with the vitrification in fiber plugs, provides good quality sex-known embryos which survive vitrification at similar rates than embryos produced with conventional unsorted sperm; also it produces good pregnancy rates after transfer of sexed embryos both fresh and after vitrification and warming.

Cryopreservation method and composition of the vitrification solution affect viability of in vitro bovine embryos

“…We observed that bovine IVP embryos survived vitrification at higher rates than freezing by the slow-controlled rate method. Previous studies have shown similar results (Nedembale et al, 2004;Kuwayama , 2007;Rios et al, 2010;Trigal et al, 2012; attributed to the brief exposure to cryoprotectant, as described by Vajta et al (1998) and Mucci et al (2006). Membrane stability of in vitro produced bovine embryos might be an important factor determining their susceptibility and viability to freezing (Imai et al, 1997).…”

“…In terms of embryo survival after thawing/ warming, vitrification of IVP embryos has been proved to be at least as efficient (Inaba et al, 2011) or better (Vajta and Nagy, 2006;Trigal et al, 2012;Vajta, 2013) than slow-rate freezing, so it is not easy to decide for one methodology over the other; each laboratory should decide which one adapts better to their conditions (Caamaño et al, 2015).…”

Parâmetros da produção in vitro de embriões da raça Sindi

“…Do mesmo modo, Trigal et al (2012) confirmaram o efeito individual do touro, assim como do tipo de sêmen sobre a taxa de blastocistos, que foi maior com o sêmen não sexado do que com o sexado.…”

Verifikasi molekuler metode sexing sperma sapi dengan kolom BSA (Bovine Serum Albumin)

“…Struktur gen SRY tersebut memiliki daerah sentral yang berkorelasi dengan HMG (homeobox) atau disebut dengan housekeeping gene (pada mamalia disebut dengan GADPH) yang memiliki daerah konservatif yang dapat membedakan antar spesies. Karakteristik SRY-HMG inilah yang biasa digunakan untuk penelitian DNA berbasis penentuan jenis kelamin (Taylor 2005 ;Choi et al 2009 ;Trigal et al 2012 ;Gokulakrishnan et al 2015).…”