Influence of Culture Medium Composition on Relative mRNA Abundances in Domestic Cat Embryos

Abstract

Contents Different culture conditions have been used to produce domestic cat embryos. As part of the in vitro procedures, the medium composition significantly affects the quality of the embryo development also. Quality assessments based on cleavage kinetics and blastomere symmetry are useful, but embryos also can differ in their relative gene expression patterns despite similar morphological characteristics. The aim of this study was to compare cat embryos produced with two different in vitro culture systems routinely used in two different laboratories [Smithsonian Conservation Biology Institute, Washington D.C., USA (SCBI) and Leibniz Institute for Zoo and Wildlife Research, Berlin, Germany (IZW)]. Specifically, relative mRNA expression patterns of critical genes for pre‐implantation embryo development were assessed in both conditions. Embryos were produced in parallel in both culture systems by IVF using frozen‐thawed ejaculated semen in the United States and fresh epididymal sperm in Germany. Success of embryo development in vitro was recorded as well as relative mRNA abundances [DNA methyltransferases 1 and 3A (DNMT1, DNMT3A), gap junction protein alpha 1 (GJA1), octamer‐binding transcription factor 4 [OCT4], insulin‐like growth factors 1 and 2 receptors (IGF1R, IGF2R), beta‐actin (ACTB)] in pools of days 4–5 morulae by semi‐quantitative RT‐PCR assay. Percentages of cleaved embryos were similar (p > 0.05) between both culture systems, regardless of the location. OCT4 mRNA abundance was higher (p < 0.05) in embryos derived in the SCBI culture system compared with those from the IZW system when epididymal sperm was used for IVF. No clear correlation between the expression pattern and the culture system could be found for all other genes. It is suggested that OCT4 expression might be affected by the media composition in some conditions and can be the indicator of a better embryo quality.

Capabilities and Challenges of Examination of Gene Expression for Quality Assessment of Domestic Cat Embryos

“…The alteration in the procedure could be responsible for the diverging results, as it was shown previously that the culture medium composition affects relative mRNA abundance in cat embryos (Hribal et al. ). It can be hypothesized that embryos produced in Ham's F10 are able to compensate negative effects caused by the sperm source better.…”

“…OCT4 expression was furthermore influenced by the culture medium composition used for the production of cat embryos (Hribal et al. ). The impact of ICSI and sperm cryopreservation, both long‐term applied methods in the domestic cat (Pope et al.…”

“…OCT4 expression was decreased in cloned blastocysts produced with frozen-thawed donor fibroblast cells compared with fresh cells or IVF embryos (Gomez et al 2008). OCT4 expression was furthermore influenced by the culture medium composition used for the production of cat embryos (Hribal et al 2012). The impact of ICSI and sperm cryopreservation, both long-term applied methods in the domestic cat (Pope et al 1998;Ringleb et al 2011), on expression of specific genes at morula stage has been examined previously (Waurich et al 2010).…”

The never‐ending search of an ideal culture system for domestic cat oocytes and embryos

“…Investigation on embryo gene expression could also contribute to understand the effects of the in vitro conditions and the specific embryo requirements during IVC. In the domestic cat, it has been proved that gene expression varies according to the embryo stages, and differs between in vitro and in vivo-derived embryos (Filliers et al, 2012), depending also on the culture medium (Hribal, Jewgenow, Braun, & Comizzoli, 2013) and on the spermatozoa status (fresh or cryopreserved; Hribal, Braun, Ringleb, & Jewgenow, 2012).…”

Baicalin increases developmental competence of mouse embryos <i>in vitro</i> by inhibiting cellular apoptosis and modulating <i>HSP70</i> and DNMT expression

“…In vitro culture of preimplantation embryos is an essential step in assisted reproductive technology (ART) for both human and animals [1, 2], which is now considered to be a part of mainstream medical practice. However, the birth rate of offspring following ART still lags behind that of their in vivo counterparts, with only a certain percentage of in vitro -cultured (IVC) embryos being capable of establishing pregnancy after their transfer into recipients [3].…”