The inhibition of adipogenesis via an in vitro assay can reduce animal use by more precisely estimating the starting dose for the acute toxic class method

Abud, Ana Paula Ressetti; Kuligovski, Crisciele; Corrêa, Natássia Caroline Resende; Moraes, Elizabeth Cunha Penna de; Caruso, Rodrigo Rêgo Barros; Schuck, Desirée Cigaran; Brohem, Carla Abdo; Dallagiovanna, Bruno; Aguiar, Alessandra Melo de

doi:10.1016/j.toxlet.2019.04.009

Cited by 4 publications

(13 citation statements)

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“…Due to their advantages of self-renewal capability and cell differentiation, stem cells can be very useful for the evaluation of toxicological mechanisms. These specific characteristics may provide additional strategies and applicable data for human toxicity predictions and to improve sensitivity [3 , 4] . In addition, it is possible to carry out tests on different biological samples to evaluate biological variability [4] , [5] , [6] , [7] .…”

Section: Methods Introductionmentioning

confidence: 99%

“…These specific characteristics may provide additional strategies and applicable data for human toxicity predictions and to improve sensitivity [3 , 4] . In addition, it is possible to carry out tests on different biological samples to evaluate biological variability [4] , [5] , [6] , [7] . Moreover, stem cells derived from tissue removed during surgery, specifically liposuction procedures [4 , 8] , are of great interest because they represent disposable material, are easy to handle, and do not require a wide variety of supplements to culture [9] .…”

Section: Methods Introductionmentioning

confidence: 99%

“…A major concern worldwide is the poor correlation between in vitro and in vivo data. After our group obtained the abovementioned results, we questioned whether the ability of stem cells to differentiate could provide a methodology that would result in more accurate data to assess toxicity in vitro [4] . We chose adipogenesis because it is a very well-known, easily quantifiable cell differentiation model; thus, adipogenesis is more economically viable than pluripotent cell models, such as cardiac differentiation.…”

Section: Additional Informationmentioning

confidence: 99%

“…This method makes it possible to use cells of human origin, and our previous data indicate that evaluating the toxic effects during the adipogenic differentiation process is more sensitive with relative viability assays, similar to traditionally used methods, such as the NRU assay [12] . We evaluated 10 test substances with a high-content imaging system, and the data provided by this test indicated that the inhibition of adipogenic differentiation predicted the toxicity of highly toxic substances more accurately than the NRU assay [4] . Thus, we suggest the use of this test as an alternative method to the use of animals that provides complementary results to the NRU assay.…”

Section: Additional Informationmentioning

confidence: 99%

“…In our previous study [4] , we developed an assay quality control system that compares negative and positive internal controls according to the quality control criterion. We evaluated three quality control criteria: the Z’ factor; strictly standardized mean difference (SSMD); and robust SSMD.…”

Section: Additional Informationmentioning

confidence: 99%

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Using inhibition of the adipogenesis of adipose-derived stem cells in vitro for toxicity prediction

et al. 2021

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In vitro stem cell models are used as alternatives to animal models and are important tools for cytotoxicity studies. Researchers can determine the effects of test substances on human cells by evaluating cell viability and differentiation. Here, we describe an in vitro model to quantify adipogenesis based on the Nile red staining of specific lipid droplets and the emission of basic lipids from human adipose tissue-derived mesenchymal stromal cells (AD-MSCs) in the presence of test substances. This assay allows for the prediction of toxicity based on the inhibition of adipogenesis in vitro in a 96-well format. The differentiation of a progenitor cell into a specialized cell, the adipocyte, is easy to monitor and quantify, making this a simple assay. The fluorescence staining of nuclei and lipid droplets is measured after 14 days of cell differentiation to determine cell number and assess cell differentiation using high-content imaging analysis, thus allowing for the identification of chemicals that impact differentiation. We also describe a protocol to assess adipocyte differentiation by fluorescence intensity using a multiplate reader. Researchers can utilize the protocol described here for many purposes to evaluate in vitro adipogenesis. With this method, it is possible to reduce the use of animals.

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