Selection of theIn VitroCulture Media Influences mRNA Expression of Hedgehog Genes,Il-6,and Important Genes regarding Reactive Oxygen Species in Single Murine Preimplantation Embryos

Author:

Pfeifer N.1,Baston-Büst D. M.1,Hirchenhain J.1,Friebe-Hoffmann U.1,Rein D. T.2,Krüssel J. S.1,Hess A. P.1

Affiliation:

1. Department of OB/GYN and REI (UniKiD), Medical Center University of Düsseldorf, Moorenstr. 5, 40225 Düsseldorf, Germany

2. St. Elisabeth Hospital Hohenlind, Department of OB/GYN, Werthmannstr. 1, 50935 Köln, Germany

Abstract

Background. The aim of this paper was to determine the influence of differentin vitroculture media on mRNA expression of Hedgehog genes,il-6,and important genes regarding reactive oxygen species in single mouse embryos.Methods. Reverse transcription of single embryos either culturedin vitrofrom day 0.5 until 3.5 (COOK’s Cleavage medium or Vitrolife’s G-1 PLUS medium) orin vivountil day 3.5post coitum. PCR was carried out forβ-actinfollowed by nested-PCR forshh, ihh, il-6, nox, gpx4, gpx1,andprdx2.Results. The number of murine blastocysts cultured in COOK medium which expressedil-6, gpx4, gpx1,andprdx2mRNA differed significantly compared to thein vivogroup. Except fornox, the mRNA profile of the Vitrolife media group embryos varied significantly from thein vivoones regarding the number of blastocysts expressing the mRNA ofshh, ihh, il-6, gpx4, gpx1andprdx2.Conclusions. The present study shows that differentin vitroculture media lead to different mRNA expression profiles during early development. Even the newly developedin vitroculture media are not able to mimic the female reproductive tract. The question of long-term consequences for children due to assisted reproduction techniques needs to be addressed in larger studies.

Publisher

Hindawi Limited

Subject

General Environmental Science,General Biochemistry, Genetics and Molecular Biology,General Medicine

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