Data di Pubblicazione:
2010
Citazione:
Different ability to form outgrowth between in vitro produced porcine embryos and parthenote / G. Pennarossa, A. Vanelli, F. Gandolfi, T.A.L. Brevini. ((Intervento presentato al 3. convegno Maternal Interactions with Gametes and Embryos tenutosi a Soustons nel 2010.
Abstract:
The establishment of porcine embryonic stem cell (pESC) lines would be a novel tool for animal biotechnology and it would represent a useful model for biomedical research as well as developmental biology studies. In the last years parthenotes received particular attention because they represent an alternative ethical source to obtain ESC lines. However little is known about the biological differences between parthenotes and embryos and further studies are needed in order to better elucidate these aspects. Here we produced in vitro fertilized (IVF) and parthenogenetic embryos and isolated ICMs in order to compare their ability to form outgrowths.
More in detail, blastocysts were subjected to immuno-surgery, using pronase 0.5% (w/v), monkey anti-porcine serum (supplied by Istituto Zooprofilattico Sperimentale Lombardia ed Emilia- Romagna) and Guinea pig complement. ICMs were isolated from lysed trophoblast cells by pipetting and plated onto inactivated STO feeder cells and outgrowth formation was monitored.
We observed that parthenogenetic ICMs generated a significantly higher number of outgrowths than IVF ones (22.16% vs 4.82%). In order to better understand this result we compared the expression levels of the cell adhesion molecules beta integrin-1and vitronectin in ICMs obtained from the two sources.
A statistically significant difference for the two genes was detected. In particular, the expression of beta integrin-1 was 5,6-fold higher in parthenotes than in IVF ICMs (ΔCt values: 68±0.5 vs 12±0.1). Similarly vitronectin displayed an up regulation 11,7-fold in parthenogenetic embryos (ΔCt values: 82±0.7 vs 7.3±0.2).
Our findings provide the first molecular explanation for the higher efficiency to generate outgrowths exhibited by parthenogenetic blastocysts, as compared to regular IVF embryos.
Tipologia IRIS:
14 - Intervento a convegno non pubblicato
Elenco autori:
G. Pennarossa, A. Vanelli, F. Gandolfi, T.A.L. Brevini
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