Ethanol, Acetaldehyde, and Estradiol Affect Growth and Differentiation of Rhesus Monkey Embryonic Stem Cells

Abstract

Background The timing of the origins of fetal alcohol syndrome have been difficult to determine, in part because of the challenge associated with in vivo studies of the peri-implantation stage of embryonic development. Because embryonic stem cells (ESCs) are derived from blastocyst stage embryos, they are used as a model for early embryo development. Methods Rhesus monkey ESC lines (ORMES-6 and -7) were treated with 0, 0.01%, 0.1%, or 1.0% ethanol, 1.0% ethanol with estradiol or 0.00025% acetaldehyde with or without estradiol for 4 weeks. Results Although control ESCs remained unchanged, abnormal morphology of ESCs in the ethanol and acetaldehyde treatment groups was observed before 2 weeks of treatment. Immunofluorescence staining of key pluripotency markers ( TRA-1- 81 and alkaline phosphatase) indicated a loss of ESC pluripotency in the 1.0% ethanol group. ORMES-7 was more sensitive to effects of ethanol than ORMES-6. Conclusions Estradiol appeared to increase sensitivity to ethanol in the ORMES-6 and -7 cell line. The morphological changes and labeling for pluripotency, proliferation and apoptosis demonstrated that ethanol affects how these early cells develop in culture, their differentiation state in particular. The effects of ethanol may be mediated in part through metabolic pathways regulating acetaldehyde formation, and while potentially accentuated by estradiol in some individuals, how remains to be determined.

Different Effects of Knockouts in ALDH2 and ACSS2 on Embryonic Stem Cell Differentiation

“…E to G ). These results are consistent with a model of EtOH stimulating stem cell differentiation (Serio et al., ; VandeVoort et al., ). The delay in differentiation observed by Arzumnayan and colleagues () may have been unique to the method of differentiation being utilized, via depletion of LIF in cell culture medium.…”

“…Context‐dependent changes in pluripotency and differentiation‐related factors have been demonstrated by using EtOH to investigate effects of stem cell differentiation (Arzumnayan et al., ; Ogony et al., ; VandeVoort et al., ; Veazey et al., ). For example, treatment of ESCs with a wide range of EtOH doses, from 0.1% to 1% (17.4 to 174 mM), for 2 weeks caused a loss of pluripotency and spontaneous differentiation, as measured by alkaline phosphatase and TRA‐1‐81 staining, despite some pluripotency‐related genes remaining highly expressed (VandeVoort et al., ). In contrast to these findings, a 48‐hour treatment with 100 mM EtOH followed by 6 days of differentiation via LIF removal caused a delay in the loss of pluripotency factor–related mRNAs, suggesting that differentiation was also being delayed by EtOH (Arzumnayan et al., ).…”

Gene expression signatures affected by alcohol-induced DNA methylomic deregulation in human embryonic stem cells

“…Specifically, we demonstrated that a 24 hr low dose (20 mM) EtOH treatment significantly reduced the pluripotency (differentiation potential) of hESCs. While to our knowledge no other hESC studies have addressed this issue, somewhat analogous observations were made with rhesus monkey ESCs, albeit at much higher EtOH concentrations and over a course of 4 weeks (VandeVoort et al, 2011). We also, for the first time, demonstrated overall increases in DNA methylation in hESCs, defined the genetic and epigenetic molecular landscapes affected by low dose EtOH exposure, and identified genome-wide hotspots that could potentially be vulnerable to FAE.…”

Ethanol Alters the Balance of Sox2, Oct4, and Nanog Expression in Distinct Subpopulations During Differentiation of Embryonic Stem Cells

“…A considerable body of literature has addressed the effects of ethanol in mouse and human ES and neural stem cells [26][27][28][29][30][31][32][33][34][35][36]. Long-term in vitro ethanol exposure as a model of binge drinking has been used to study the molecular aspects of FAS.…”