Use of Video Bioinformatics Tools in Stem Cell Toxicology

Talbot, Prue; Nieden, Nicole I. zur; Lin, Sabrina; Martinez, Ivann Kenneth Carvajal; Guan, Ben Bo; Bhanu, Bir

doi:10.1002/9781118856017.ch21

Cited by 22 publications

(30 citation statements)

References 77 publications

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“…Images were obtained every 2 h for 48 h, then analyzed using CL Quant software (DR Vision, Seattle, WA) to track dynamic cell processes, such as proliferation and death (Lin et al, 2010a, 2010b; Guan et al, 2014; Talbot et al, 2014). …”

Section: Methodsmentioning

confidence: 99%

Thirdhand smoke: Chemical dynamics, cytotoxicity, and genotoxicity in outdoor and indoor environments

Bahl

Shim

Jacob

et al. 2016

Toxicology in Vitro

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We tested the toxicity of thirdhand smoke (THS) using two controlled laboratory exposure scenarios and low levels of THS. One exposure modeled THS in a car parked outdoors, while the second modeled THS in a room without sunlight. The fabrics were exposed to cigarette smoke and then extracted in culture medium. Concentrations of nicotine, nicotine related alkaloids, and tobacco specific nitrosamines (TSNAs) were determined in fresh and aged extracts. The concentration of TSNAs increased with aging in the indoor experiment. THS extracts were used for cytotoxicity testing using mouse neural stem cells (mNSC), human dermal fibroblasts (hDF) and human palatal mesenchyme cells (hPM). Extracts from the car experiment inhibited mNSC proliferation in a live cell imaging assay and induced single strand DNA breaks in mNSC and hDF. In the indoor experiment, THS extracts made with medium containing serum proteins were significantly more toxic than extracts made with basal medium, and mNSC and hPM were more sensitive than hDF. These data indicate that: (1) aging of THS chemical differs on different fabrics and differs with and without sunlight; (2) very few cigarettes are sufficient to produce a toxic THS residue; and (3) protein enhances the efficiency of extraction of cytotoxic chemicals.

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Section: Methodsmentioning

confidence: 99%

Thirdhand smoke: Chemical dynamics, cytotoxicity, and genotoxicity in outdoor and indoor environments

Bahl

Shim

Jacob

et al. 2016

Toxicology in Vitro

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“…Because stem cells are present throughout life, their risk of accumulating damage is high. Stem cells are also often more sensitive to stress than differentiated cells (Bahl et al., 2012, Behar et al., 2014), making them excellent models for evaluating safe limits of exposure to potential toxicants (Betts, 2010, Liu et al., 2017, Talbot et al., 2014, Talbot and Lin, 2011, Yin et al., 2015).…”

Section: Introductionmentioning

confidence: 99%

Mitochondrial Stress Response in Neural Stem Cells Exposed to Electronic Cigarettes

et al. 2019

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Summary Stem cells provide a sensitive model to study exposure to toxicants, such as cigarette smoke. Electronic cigarettes (ECs) are popular nicotine delivery devices, often targeted to youth and pregnant mothers. However, little is known about how chemicals in ECs might affect neural stem cells, and in particular their mitochondria, organelles that maintain cell functionality and health. Here we show that the mechanism underlying EC-induced stem cell toxicity is stress-induced mitochondrial hyperfusion (SIMH), a transient survival response accompanied by increased mitochondrial oxidative stress. We identify SIMH as a survival response to nicotine, now widely available in EC refill fluids and in pure form for do-it-yourself EC products. These observed mitochondrial alterations combined with autophagy dysfunction to clear damaged mitochondria could lead to faulty stem cell populations, accelerate cellular aging, and lead to acquired mitochondriopathies. Any nicotine-containing product may likewise stress stem cells with long-term repercussions for users and passively exposed individuals. Video Abstract

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“…A Nikon BioStation IM, which combines an incubator, microscope, and cooled CCD camera, was used to collect time-lapse video data ( Weng et al, 2015 ; Talbot et al, 2014 ; Lin et al, 2015 ). To create time-lapse videos, human prostate cancer cells, mEF, mESC, and hESC were plated on 35 mm culture dishes and incubated in a BioStation IM.…”

Section: Methodsmentioning

confidence: 99%

Dynamic blebbing: A bottleneck to human embryonic stem cell culture that can be overcome by Laminin-Integrin signaling

2018

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This study characterizes dynamic and apoptotic blebbing in human embryonic stem cells (hESC), identifies dynamic blebbing as a bottleneck to successful cell attachment during passaging, and demonstrates that dynamic blebbing can be rapidly stopped by plating cells on recombinant human laminin. In freshly plated hESC, dynamic and apoptotic blebbing differed in time of occurrence, bleb retraction rate, mitochondrial membrane potential, and caspase 3&7 activation. While dynamic blebbing can be controlled with drugs that inhibit myosin II, these methods have off-target effects and are not suitable for clinical applications. Recombinant human laminin-521 or addition of laminin-111 to Matrigel provided a safe method to drastically decrease dynamic blebbing and improve cell attachment with proteins normally found in the inner cell mass. Inhibition of focal adhesion kinase, which is activated by binding of integrins to laminin, prolonged dynamic blebbing and inhibited attachment. These data show that hESC bind rapidly to laminins through an integrin, which activates focal adhesion kinase that in turn downregulates dynamic blebbing. Laminins enabled hESC to rapidly attach during passaging, improved plating efficiency, enabled passaging of single pluripotent stem cells, and avoided use of inhibitors that have non-specific off-target effects. These data provide a strategy for improving hESC culture using biologically safe recombinant human proteins.

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Use of Video Bioinformatics Tools in Stem Cell Toxicology

Cited by 22 publications

References 77 publications

Thirdhand smoke: Chemical dynamics, cytotoxicity, and genotoxicity in outdoor and indoor environments

Thirdhand smoke: Chemical dynamics, cytotoxicity, and genotoxicity in outdoor and indoor environments

Mitochondrial Stress Response in Neural Stem Cells Exposed to Electronic Cigarettes

Dynamic blebbing: A bottleneck to human embryonic stem cell culture that can be overcome by Laminin-Integrin signaling

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