Direct comparison of distinct naive pluripotent states in human embryonic stem cells

Stem cells & developmental biolog

A mRNA landscape of bovine embryos after standard and MAPK-inhibited culture conditions: a comparative analysis.

BACKGROUND: Genes and signalling pathways involved in pluripotency have been studied extensively in mouse and human pre-implantation embryos and embryonic stem (ES) cells. The unsuccessful attempts to generate ES cell lines from other species including cattle suggests that other genes and pathways are involved in maintaining pluripotency in these species. To investigate which genes are involved in bovine pluripotency, expression profiles were generated from morula, blastocyst, trophectoderm and inner cell mass (ICM) samples using microarray analysis. As MAPK inhibition can increase the NANOG/GATA6 ratio in the inner cell mass, additionally blastocysts were cultured in the presence of a MAPK inhibitor and changes in gene expression in the inner cell mass were analysed. RESULTS: Between morula and blastocyst 3,774 genes were differentially expressed and the largest differences were found in blastocyst up-regulated genes. Gene ontology (GO) analysis shows lipid metabolic process as the term most enriched with genes expressed at higher levels in blastocysts. Genes with higher expression levels in morulae were enriched in the RNA processing GO term. Of the 497 differentially expressed genes comparing ICM and TE, the expression of NANOG, SOX2 and POU5F1 was increased in the ICM confirming their evolutionary preserved role in pluripotency. Several genes implicated to be involved in differentiation or fate determination were also expressed at higher levels in the ICM. Genes expressed at higher levels in the ICM were enriched in the RNA splicing and regulation of gene expression GO term. Although NANOG expression was elevated upon MAPK inhibition, SOX2 and POU5F1 expression showed little increase. Expression of other genes in the MAPK pathway including DUSP4 and SPRY4, or influenced by MAPK inhibition such as IFNT, was down-regulated. CONCLUSION: The data obtained from the microarray studies provide further insight in gene expression during bovine embryonic development. They show an expression profile in pluripotent cells that indicates a pluripotent, epiblast-like state. The inability to culture ICM cells as stem cells in the presence of an inhibitor of MAPK activity together with the reported data indicates that MAPK inhibition alone is not sufficient to maintain a pluripotent character in bovine cells

Adaptive optics assisted Fourier domain OCT with balanced detection

Two factors are of importance to optical coherence tomography (OCT), resolution and sensitivity. Adaptive optics improves the resolution of a system by correcting for aberrations causing distortions in the wave-front. Balanced detection has been used in time domain OCT systems by removing excess photon noise, however it has not been used in Fourier domain systems, as the cameras used in the spectrometers saturated before excess photon noise becomes a problem. Advances in camera technology mean that this is no longer the case and balanced detection can now be used to improve the signal to noise ratio in a Fourier domain (FD) OCT system. An FD-OCT system, enhanced with adaptive optics, is presented and is used to show the improvement that balanced detection can provide. The signal to noise ratios of single camera detection and balanced detection are assessed and in-vivo retinal images are acquired to demonstrate better image quality when using balance detection

Human blastocyst outgrowths recapitulate primordial germ cell specification events

Stem cells & developmental biolog

Media 1: Real-time microscopic phase-shifting profilometry

Originally published in Applied Optics on 20 May 2015 (ao-54-15-4953

The TGF beta pathway plays a pivotal role in early embryo lineage segregation in both mouse and human

Stem cells & developmental biolog

Dose-dependent effect of inhibition of the transforming growth factor beta pathway during early mouse lineage segregation

Stem cells & developmental biolog

The Influence of Early Embryo Traits on Human Embryonic Stem Cell Derivation Efficiency

Despite its prognostic value in in vitro fertilization, early embryo morphology is not reported on in the derivation of human embryonic stem cell (hESC) lines. Standard hESC derivation does rely on blastocyst development and its efficiency is highly correlated to inner cell mass (ICM) quality. Poor-quality embryos (PQEs) donated for hESC derivation may have a range of cleavage-stage abnormalities that are known to compromise further development. This study was implemented to determine whether specific PQEs traits influence the efficiency of good-quality ICMs to derive new hESC lines. We found that although the types of PQEs investigated were all able to make blastocysts with good-quality ICMs, the ICMs were unequal in their ability to derive hESCs. Good-quality ICMs from embryos with multiple poor-quality traits were unable to generate hESC lines, in contrast to good-quality ICMs from embryos with a single poor-quality trait. In addition, our data suggest a direct correlation between the number of ICM cells present in the blastocyst and its capacity to derive new hESC lines. This study is the first to demonstrate that ICM quality alone is an incomplete indicator of hESC derivation and that application of in vitro fertilization-based early embryo scoring can help predict hESC derivation efficiency. Experiments aiming to quantify, improve upon, or compare hESC derivation efficiency should thus take into consideration early embryo morphology scoring for the comparison of groups with equal developmental competence.Stem cells & developmental biolog

A educação científica pelos artefatos tecnológicos

Este trabalho propõe abordar os objetos industrializados como instrumentos pedagógicos da educação científica, colocando em evidência as relações entre conhecimento, inventividade, o imaginário e a realidade científica e tecnológica da sociedade contemporânea