Molecular effect of ethanol during neural differentiation of human embryonic stem cells in vitro

Abstract

Potential teratogenic effects of alcohol on fetal development have been documented. Especially studies have demonstrated deleterious effect of ethanol exposure on neuronal development in animal models and on the maintenance and differentiation of neuronal precursor cells derived from stem cells. To better understand molecular effect of alcohol on the process of neural differentiation, we have performed gene expression microarray analysis on human embryonic stem cells being directed to neural rosettes and neural precursor cells in the presence of ethanol treatment. Here we provide detailed experimental methods, analysis and information associated with our data deposited into Gene Expression Omnibus (GEO) under GSE56906. Our data provide scientific insight on potential molecular effects of fetal alcohol exposure on neural differentiation of early embryo development.

Time- and dose-dependent effects of ethanol on mouse embryonic stem cells

“…Other differentiation markers were shown to be dysregulated by ethanol in Rhesus monkey ES cells [40]. Indeed, altered expression of numerous genes and/or proteins as well as metabolites have been reported in human and mouse ES cells, neural stem cells, induced pluripotent stem cells, pluripotent carcinoma cells, or cells derived from those cells [36][37][38][39][40][41][42][43][44][45][46][47][48][49][50]. The concentrations of ethanol that were most commonly used in these studies were in the range of 0.1% to 0.5% with the justification that 0.12% is equivalent to DUI levels and 0.29%-0.5% falls within the levels measured in alcoholics [51].…”

Alcohol-Induced Molecular Dysregulation in Human Embryonic Stem Cell-Derived Neural Precursor Cells

“…Human embryonic stem cells (H1 and H9 lines) obtained through license agreement with WiCell Research Institute (Madison, WI) were cultured on mouse embryonic fibroblast (MEF) feeder layer and were transferred to mTeSR1 serum free human embryonic stem cell (hESC) culture system (STEMCELL Technologies Inc., Vancouver, Canada). Neural differentiation of hESCs was performed by using STEMdiff Neural System (STEMCELL Technologies Inc., Vancouver, Canada) according to the manufacturer’s instruction as described in our previous publications [ 23 , 24 ]. After 7 day differentiation, morphological assessment and scoring of neural rosettes were done to ensure 50% or more of the area of each aggregate was filled with neural rosettes (as shown in Fig 1 ).…”

“…RNA isolation and microarray analysis was done as described [ 20 , 24 ]. Briefly, the neural aggregates at D10 for the formation of rosettes and at D15 (5 days after re-plating the rosette clusters for the expansion of neural precursor cells) were used for total RNA isolation by using RNeasy mini kit (Qiagen, Valencia, CA).…”

A genomics-based framework for identifying biomarkers of human neurodevelopmental toxicity

“…Secondary filters eliminated studies that used genetically modified lines, non-neural-related differentiation routes, and non-human cell lines. In total, we identified four unique hESC neural differentiation datasets [36][37][38][39]. We added a transcriptomic dataset that was generated in our laboratory that compared the UCSF4 hESC line in a pluripotent state and their NPC derivatives [19].…”