Cell-based assay for the detection of chemically induced cellular stress by immortalized untransformed transgenic hepatocytes

Sacco, Maria Grazia; Amicone, Laura; Catò, Enrica Mira; Filippini, Daniela; Vezzoni, Paolo; Tripodi, Marco

doi:10.1186/1472-6750-4-5

Cited by 13 publications

(1 citation statement)

References 22 publications

(33 reference statements)

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“…HepE14 are hepatocytes displaying a differentiated phenotype and a coherent gene expression profile. They have been used in a variety of studies of hepatocyte physiology being able to express a wide range of liver functions and products 34,54–58 . HepE14 cells were grown at 37 °C, in a humidified atmosphere with 5% CO2 on collagen I (Collagen I, Rat Tail; Gibco-Life Technologies) coated dishes in RPMI-1640 medium (Gibco-Life Technologies), supplemented with 10% FBS, 2 mM glutamine (Gibco-Life Technologies), 50 ng/ml EGF, 30 ng/ml IGF II (PeproTech), 10 µg/ml insulin (Roche) and antibiotics.…”

Section: Methodsmentioning

confidence: 99%

YAP integrates the regulatory Snail/HNF4α circuitry controlling epithelial/hepatocyte differentiation

et al. 2019

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Yes-associated protein (YAP) is a transcriptional co-factor involved in many cell processes, including development, proliferation, stemness, differentiation, and tumorigenesis. It has been described as a sensor of mechanical and biochemical stimuli that enables cells to integrate environmental signals. Although in the liver the correlation between extracellular matrix elasticity (greatly increased in the most of chronic hepatic diseases), differentiation/functional state of parenchymal cells and subcellular localization/activation of YAP has been previously reported, its role as regulator of the hepatocyte differentiation remains to be clarified. The aim of this study was to evaluate the role of YAP in the regulation of epithelial/hepatocyte differentiation and to clarify how a transducer of general stimuli can integrate tissue-specific molecular mechanisms determining specific cell outcomes. By means of YAP silencing and overexpression we demonstrated that YAP has a functional role in the repression of epithelial/hepatocyte differentiation by inversely modulating the expression of Snail (master regulator of the epithelial-to-mesenchymal transition and liver stemness) and HNF4α (master regulator of hepatocyte differentiation) at transcriptional level, through the direct occupancy of their promoters. Furthermore, we found that Snail, in turn, is able to positively control YAP expression influencing protein level and subcellular localization and that HNF4α stably represses YAP transcription in differentiated hepatocytes both in cell culture and in adult liver. Overall, our data indicate YAP as a new member of the HNF4/Snail epistatic molecular circuitry previously demonstrated to control liver cell state. In this model, the dynamic balance between three main transcriptional regulators, that are able to control reciprocally their expression/activity, is responsible for the induction/maintenance of different liver cell differentiation states and its modulation could be the aim of therapeutic protocols for several chronic liver diseases.

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Section: Methodsmentioning

confidence: 99%

YAP integrates the regulatory Snail/HNF4α circuitry controlling epithelial/hepatocyte differentiation

et al. 2019

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Overview of Stem and Artificial Cells

Soto-Gutiérrez

Navarro-Alvarez

Rivas‐Carrillo

et al. 2010

Pharmaceutical Sciences Encyclopedia

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Mammalian Cell-Based Sensor System

Banerjee

Franz

Bhunia

2010

Whole Cell Sensing Systems I

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Use of living cells or cellular components in biosensors is receiving increased attention and opens a whole new area of functional diagnostics. The term "mammalian cell-based biosensor" is designated to biosensors utilizing mammalian cells as the biorecognition element. Cell-based assays, such as high-throughput screening (HTS) or cytotoxicity testing, have already emerged as dependable and promising approaches to measure the functionality or toxicity of a compound (in case of HTS); or to probe the presence of pathogenic or toxigenic entities in clinical, environmental, or food samples. External stimuli or changes in cellular microenvironment sometimes perturb the "normal" physiological activities of mammalian cells, thus allowing CBBs to screen, monitor, and measure the analyte-induced changes. The advantage of CBBs is that they can report the presence or absence of active components, such as live pathogens or active toxins. In some cases, mammalian cells or plasma membranes are used as electrical capacitors and cell-cell and cell-substrate contact is measured via conductivity or electrical impedance. In addition, cytopathogenicity or cytotoxicity induced by pathogens or toxins resulting in apoptosis or necrosis could be measured via optical devices using fluorescence or luminescence. This chapter focuses mainly on the type and applications of different mammalian cell-based sensor systems.

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Cell-based assay for the detection of chemically induced cellular stress by immortalized untransformed transgenic hepatocytes

Cited by 13 publications

References 22 publications

YAP integrates the regulatory Snail/HNF4α circuitry controlling epithelial/hepatocyte differentiation

YAP integrates the regulatory Snail/HNF4α circuitry controlling epithelial/hepatocyte differentiation

Overview of Stem and Artificial Cells

Mammalian Cell-Based Sensor System

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