Selection of the in vitro culture media influences mRNA expression of Hedgehog genes, Il-6, and important genes regarding reactive oxygen species in single murine preimplantation embryos.
BACKGROUND
The aim of this paper was to determine the influence of different in vitro culture 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 cultured in vitro from day 0.5 until 3.5 (COOK's Cleavage medium or Vitrolife's G-1 PLUS medium) or in vivo until day 3.5 post coitum. PCR was carried out for β-actin followed by nested-PCR for shh, ihh, il-6, nox, gpx4, gpx1, and prdx2.


RESULTS
The number of murine blastocysts cultured in COOK medium which expressed il-6, gpx4, gpx1, and prdx2 mRNA differed significantly compared to the in vivo group. Except for nox, the mRNA profile of the Vitrolife media group embryos varied significantly from the in vivo ones regarding the number of blastocysts expressing the mRNA of shh, ihh, il-6, gpx4, gpx1 and prdx2.


CONCLUSIONS
The present study shows that different in vitro culture media lead to different mRNA expression profiles during early development. Even the newly developed in vitro culture 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.
Department of OB/GYN and REI (UniKiD), Medical Center University of Düsseldorf, Moorenstr. 5, 40225 Düsseldorf, Germany.
N
N
Pfeifer
D M
DM
Baston-Büst
J
J
Hirchenhain
U
U
Friebe-Hoffmann
D T
DT
Rein
J S
JS
Krüssel
A P
AP
Hess
D000818:Animals; D001483:Base Sequence; D001755:Blastocyst; D003470:Culture Media; D017931:DNA Primers; D005260:Female; D053823:Hedgehog Proteins / Q000235:genetics*; D066298:In Vitro Techniques; D015850:Interleukin-6 / Q000235:genetics*; D051379:Mice; D012333:RNA, Messenger / Q000235:genetics*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D003470:Culture Media; D017931:DNA Primers; D053823:Hedgehog Proteins; D015850:Interleukin-6; D012333:RNA, Messenger; D017382:Reactive Oxygen Species