Reproducibility of adipogenic responses to metabolism disrupting chemicals in the 3T3-L1 pre-adipocyte model system: An interlaboratory study

Nonylphenol Polyethoxylates Enhance Adipose Deposition in Developmentally Exposed Zebrafish

“…3T3-L1 cells are routinely used to assess putative adipogenic activity [52,53,64,65]. However, we recently demonstrated poor inter-laboratory reproducibility in chemical responses for the 3T3-L1 cell model [37]. Given these results and the poor maintenance of this cell line over the last several decades (cell providers report very different abilities to differentiate their stock of these cells) [66][67][68], it is important to validate robust and translational models that have more direct relevance to human health outcomes.…”

“…The Zenbio hMSCs, in contrast, had no significant effects on either proliferation or cytotoxicity at any of the tested concentrations. We previously reported marked differences in the proliferation measure based on both cell models and cell source used and the brand of tissue culture plate [37,59], suggesting that this may be a more variable measure than triglyceride accumulation. Of note, the 3T3-L1 cells and both hMSC models were differentiated using distinct media formulations specific to hMSC cell line providers (as noted in methods), timelines (Zenbio cells = 14 days; Lonza cells = 21 days; manufacturer-recommended), and differentiation procedures (Zenbio = 7 days differentiation, 7 days maintenance; Lonza = 3 days differentiation, 3 days maintenance, repeated three times) which may have contributed to some of these disparate effects.…”

“…When evaluating the length of the NPEO ethoxylate chain lengths, we reported increasing adipogenic activity with increasing ethoxylate chain length up to an average length of 6, and then decreasing activity with further increases in chain length up to an average of 20 ethoxylate chains. While these results suggest robust adipogenic effects, we also recently published research demonstrating poor inter-laboratory reproducibility in chemical responses for the 3T3-L1 cell model [37], suggesting a need to substantiate this testing in more human relevant in vitro models. Moreover, we still have limited understanding of these effects in whole organisms, particularly for varying ethoxylate chain lengths.…”

The Mixture of Bisphenol-A and Its Substitutes Bisphenol-S and Bisphenol-F Exerts Obesogenic Activity on Human Adipose-Derived Stem Cells

“…It is necessary to implement standardised adipogenic differentiation protocols to improve the consistency of results. Kassotis et al recently published a comprehensive assessment of inter-laboratory reproducibility, analysing factors contributing to the variability in responses to these chemical contaminants [75]. Human mesenchymal stem cell and pre-adipocyte models require further validation to improve the translation of this knowledge to human metabolic health.…”

Exposure to Low Doses of Oxybenzone During Perinatal Development Alters Mammary Gland Stroma in Female Mice

“…Oxybenzone is a well-recognized ER-agonist ( Watanabe et al, 2015 ; Balázs et al, 2016 ) and other estrogenic chemicals have distinct effects on adipocytes isolated from different parts of the body; for example, the effects of bisphenols on the transcriptome of omental fat cells differs significantly from their effects on the transcriptome of subcutaneous adipose cells ( Peshdary et al, 2020 ). A recent interlaboratory study used the 3T3-L1 pre-adipocyte model cell line and found that triglyceride accumulation was induced by several EDCs ( Kassotis et al, 2021 ). Future studies using both cell cultures and animal models would enable us to evaluate the mechanisms that may lead to disruption of the adipose tissue in oxybenzone-treated animals.…”