scholarly journals Comparative Proteomic Characterization of 4 Human Liver-Derived Single Cell Culture Models Reveals Significant Variation in the Capacity for Drug Disposition, Bioactivation, and Detoxication

2015 ◽  
Vol 147 (2) ◽  
pp. 412-424 ◽  
Author(s):  
Rowena L. C. Sison-Young ◽  
Dimitra Mitsa ◽  
Rosalind E. Jenkins ◽  
David Mottram ◽  
Eliane Alexandre ◽  
...  
Keyword(s):  
Cell Culture ◽  
Single Cell ◽  
Significant Variation ◽  
Human Liver ◽  
Drug Disposition ◽  
Culture Models ◽  
Single Cell Culture ◽  
Cell Culture Models

scholarly journals Electron microscopy characterization of minerals formed in vitro by human bone cells and vascular smooth muscle cells

2019 ◽  
Author(s):  
Elena Tsolaki ◽  
Louis Didierlaurent ◽  
Eike Müller ◽  
Markus Rottmar ◽  
Najma Latif ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Cell Culture ◽  
Smooth Muscle ◽  
Soft Tissue ◽  
Vascular Smooth Muscle Cells ◽  
Bone Cells ◽  
Culture Models ◽  
Cell Culture Models

AbstractSoft tissue mineralization has been found to be a major component of diseases such as aortic valve stenosis and rheumatic heart disease. Cardiovascular mineralization has been suggested to follow mechanisms similar to those of bone formation with several cell culture models been developed over the years to provide mechanistic insights. These cell models have been characterized by a wide range of biochemical and molecular methods, which identified the presence of osteogenic markers and bone-like cells. However, there is a surprisingly small number of studies where the mineral formed in these cell culture models has been characterized by physico-chemical methods, and even fewer studies have compared this mineral to the one produced by bone cells in cultures. Here we investigated the morphology and composition of the minerals formed in cell cultures of vascular smooth muscle cells and bone cells. Electron microscopy and traditional cell mineralization assays were applied, revealing that vascular cells are indeed able to form calcified nodules of elemental composition similar to bone, however with different morphology. Comparison of morphologies of the two minerals to that found in cardiovascular tissue shows that some of tissue calcification resembles the calcified fibers produced by bone cells in vitro. These results suggest that the characterization of the mineral is of utmost importance and its morphology and chemical properties can contribute an important piece of information in the comprehensive analysis of soft tissue mineralization mechanisms, both in in vitro cell culture as well as in clinical samples.

scholarly journals Molecular cloning, epigenetic regulation, and functional characterization of Prkd1 gene promoter in dopaminergic cell culture models of Parkinson's disease

2015 ◽  
Vol 135 (2) ◽  
pp. 402-415 ◽  
Author(s):  
Muhammet Ay ◽  
Huajun Jin ◽  
Dilshan S. Harischandra ◽  
Arunkumar Asaithambi ◽  
Arthi Kanthasamy ◽  
...  
Keyword(s):  
Parkinson’S Disease ◽  
Cell Culture ◽  
Molecular Cloning ◽  
Epigenetic Regulation ◽  
Functional Characterization ◽  
Gene Promoter ◽  
Dopaminergic Cell ◽  
Culture Models ◽  
Cell Culture Models

Validation of in vitro cell culture models of the blood–brain barrier: Tightness characterization of two promising cell lines

2008 ◽  
Vol 97 (12) ◽  
pp. 5158-5175 ◽  
Author(s):  
Winfried Neuhaus ◽  
Verena E. Plattner ◽  
Michael Wirth ◽  
Bettina Germann ◽  
Bodo Lachmann ◽  
...  
Keyword(s):  
Cell Culture ◽  
Blood Brain Barrier ◽  
Cell Lines ◽  
Brain Barrier ◽  
In Vitro Cell Culture ◽  
Blood Brain ◽  
Culture Models ◽  
Cell Culture Models

scholarly journals Factors to consider when interrogating 3D culture models with plate readers or automated microscopes

2021 ◽  
Author(s):  
Terry Riss ◽  
O. Joseph Trask
Keyword(s):  
Cell Culture ◽  
Three Dimensional ◽  
3D Culture ◽  
Fluorescent Imaging ◽  
Culture Models ◽  
Assay Methods ◽  
Diffusion Rates ◽  
Cell Culture Models ◽  
Dimensional Cell ◽  
Cells Cultured

AbstractAlong with the increased use of more physiologically relevant three-dimensional cell culture models comes the responsibility of researchers to validate new assay methods that measure events in structures that are physically larger and more complex compared to monolayers of cells. It should not be assumed that assays designed using monolayers of cells will work for cells cultured as larger three-dimensional masses. The size and barriers for penetration of molecules through the layers of cells result in a different microenvironment for the cells in the outer layer compared to the center of three-dimensional structures. Diffusion rates for nutrients and oxygen may limit metabolic activity which is often measured as a marker for cell viability. For assays that lyse cells, the penetration of reagents to achieve uniform cell lysis must be considered. For live cell fluorescent imaging assays, the diffusion of fluorescent probes and penetration of photons of light for probe excitation and fluorescent emission must be considered. This review will provide an overview of factors to consider when implementing assays to interrogate three dimensional cell culture models.

scholarly journals Turnover of the Active Fraction of IRS1 Involves Raptor-mTOR- and S6K1-Dependent Serine Phosphorylation in Cell Culture Models of Tuberous Sclerosis

2006 ◽  
Vol 26 (17) ◽  
pp. 6425-6434 ◽  
Author(s):  
O. Jameel Shah ◽  
Tony Hunter
Keyword(s):  
Cell Culture ◽  
Tuberous Sclerosis ◽  
High Speed ◽  
Serine Phosphorylation ◽  
Feedback Signal ◽  
Insulin Receptor Substrates ◽  
Igf I ◽  
Culture Models ◽  
Cell Culture Models

ABSTRACT The TSC1-TSC2/Rheb/Raptor-mTOR/S6K1 cell growth cassette has recently been shown to regulate cell autonomous insulin and insulin-like growth factor I (IGF-I) sensitivity by transducing a negative feedback signal that targets insulin receptor substrates 1 and 2 (IRS1 and -2). Using two cell culture models of the familial hamartoma syndrome, tuberous sclerosis, we show here that Raptor-mTOR and S6K1 are required for phosphorylation of IRS1 at a subset of serine residues frequently associated with insulin resistance, including S307, S312, S527, S616, and S636 (of human IRS1). Using loss- and gain-of-function S6K1 constructs, we demonstrate a requirement for the catalytic activity of S6K1 in both direct and indirect regulation of IRS1 serine phosphorylation. S6K1 phosphorylates IRS1 in vitro on multiple residues showing strong preference for RXRXXS/T over S/T,P sites. IRS1 is preferentially depleted from the high-speed pellet fraction in TSC1/2-deficient mouse embryo fibroblasts or in HEK293/293T cells overexpressing Rheb. These studies suggest that, through serine phosphorylation, Raptor-mTOR and S6K1 cell autonomously promote the depletion of IRS1 from specific intracellular pools in pathological states of insulin and IGF-I resistance and thus potentially in lesions associated with tuberous sclerosis.

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