In vitro drug screening using a mouse stem cell-based model
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ABSTRACT:
PROVIDER: PRJEB51108 | ENA |
REPOSITORIES: ENA
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Project description:time-course experiment with embryoid bodies of CGR8 mouse embryonic stem cells ; in the whole time-series RNA from 0 days old embryoid bodies were hybridized against RNA from 3 days and 10 days old embryoid bodies Keywords = embryoid bodies Keywords = mouse Keywords = time-course Keywords = oligonucleotide array Keywords: time-course
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Project description:Progenitor cells require coordinated expression of lineage-specific genes to regulate differentiation into daughter cell types. Hopx labels cardiac progenitors that are commited to the cardiac myocyte lineage. Hopx-deficiency leads to thin myocardium in approximately mid-gestation lethality in approximately 50% of embryos (secondary to thin myocardium and presumed cardiac rupture). Hopx-/- EBs display impaired myogenesis during cardiac differentiation. ChIP-seq and RNA expression analysis suggests that Hopx down regulates Wnt signaling by directly occupying and repressing wnt ligand genes. Analysis of embryoid bodies on day 8 of cardiac differentiation. RNA was made of from 1. Hopx +/- embryoid bodies, 2. Hopx -/- embryoid bodies, Embryonic stem cell lines were derived from littermate mouse blastocysts. Results provide insight into gene programs regulated by Hopx in cardiac development.
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Project description:A high-throughput platform for assessing the activity of synthetic or natural com-pounds on the motility and development of Haemonchus contortus larvae has been estab-lished for identifying new anthelmintic compounds active against strongylid nema-todes. This study evaluated the impact of serum supplementation on larval develop-ment, motility and survival in vitro and its implications for phenotypic compound screening. Of five blood components assessed, 7.5% sheep serum significantly enhanced larval development, motility and survival compared with the original medium (LB*), leading to the formulation of an improved medium (LBS*). Proteomic analysis revealed marked differences in protein expression in larvae cultured in LBS* versus LB*, includ-ing molecules associated with structural integrity and metabolic processes. The pheno-typic screening of 240 compounds (“Global Priority Box” from Medicines Malaria Ven-ture) using LBS* yielded results distinct from those in LB*, highlighting the effect of culture conditions on screening assessments. These findings indicate/emphasise the critical need to evaluate and optimise culture media for physiologically conditions in screening platforms, improving the reliability of anthelmintic discovery.
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