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Characterization of Adipogenic Chemicals in Three Different Cell Culture Systems: Implications for Reproducibility Based on Cell Source and Handling

机译:三种不同细胞培养系统中成脂化学品的表征:对基于细胞来源和处理方式的再现性的暗示

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摘要

The potential for chemical exposures to exacerbate the development and/or prevalence of metabolic disorders, such as obesity, is currently of great societal concern. Various in vitro assays are available to assess adipocyte differentiation, though little work has been done to standardize protocols and compare models effectively. This study compares several adipogenic cell culture systems under a variety of conditions to assess variability in responses. Two sources of 3T3-L1 preadipocytes as well as OP9 preadipocytes were assessed for cell proliferation and triglyceride accumulation following different induction periods and using various tissue culture plates. Both cell line and cell source had a significant impact on potencies and efficacies of adipogenic chemicals. Gene expression analyses suggested that differential expression of nuclear receptors involved in adipogenesis underlie the differences between OP9 and 3T3-L1 cells; however, there were also differences based on 3T3-L1 cell source. Induction period modulated potency and efficacy of response depending on cell line and test chemical, and large variations were observed in triglyceride accumulation and cell proliferation between brands of tissue culture plates. Our results suggest that the selection of a cell system and differentiation protocol significantly impacts the detection of adipogenic chemicals, and therefore, influences reproducibility of these studies.
机译:当前,化学暴露加剧诸如肥胖症等代谢性疾病的发展和/或流行的潜力在社会上引起极大关注。可以进行各种体外测定来评估脂肪细胞的分化,尽管在标准化协议和有效比较模型方面所做的工作很少。这项研究在各种条件下比较了几种成脂细胞培养系统,以评估反应的变异性。在不同的诱导期和使用各种组织培养板后,评估了3T3-L1前脂肪细胞和OP9前脂肪细胞的两种来源的细胞增殖和甘油三酸酯积累。细胞系和细胞来源均对成脂化学品的效能和功效产生重大影响。基因表达分析表明,参与脂肪形成的核受体的差异表达是OP9和3T3-L1细胞之间差异的基础。但是,基于3T3-L1细胞来源也存在差异。诱导期根据细胞系和测试化学物质调节反应的效价和功效,并且在组织培养板品牌之间的甘油三酸酯积累和细胞增殖中观察到较大的差异。我们的结果表明,细胞系统和分化方案的选择会显着影响成脂化学物质的检测,因此会影响这些研究的可重复性。

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