A Three-Dimensional Lung Cell Model to Leptospira Virulence Investigations

2022 ◽  
Vol 79 (2) ◽  
Author(s):  
Camila L. Campos ◽  
Luciana R. Gomes ◽  
Ambart E. Covarrubias ◽  
Ellen E. Kato ◽  
Gisele G. Souza ◽  
...  
Keyword(s):  
Cell Model ◽  
Three Dimensional

scholarly journals An optimised GAS-pharyngeal cell biofilm model

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Heema K. N. Vyas ◽  
Jason D. McArthur ◽  
Martina L. Sanderson-Smith
Keyword(s):  
Crystal Violet ◽  
Biofilm Formation ◽  
Cell Model ◽  
Matrix Material ◽  
Three Dimensional ◽  
Growth Period ◽  
Optimal Growth ◽  
Abiotic Surfaces

AbstractGroup A Streptococcus (GAS) causes 700 million infections and accounts for half a million deaths per year. Biofilm formation has been implicated in both pharyngeal and dermal GAS infections. In vitro, plate-based assays have shown that several GAS M-types form biofilms, and multiple GAS virulence factors have been linked to biofilm formation. Although the contributions of these plate-based studies have been valuable, most have failed to mimic the host environment, with many studies utilising abiotic surfaces. GAS is a human specific pathogen, and colonisation and subsequent biofilm formation is likely facilitated by distinct interactions with host tissue surfaces. As such, a host cell-GAS model has been optimised to support and grow GAS biofilms of a variety of GAS M-types. Improvements and adjustments to the crystal violet biofilm biomass assay have also been tailored to reproducibly detect delicate GAS biofilms. We propose 72 h as an optimal growth period for yielding detectable biofilm biomass. GAS biofilms formed are robust and durable, and can be reproducibly assessed via staining/washing intensive assays such as crystal violet with the aid of methanol fixation prior to staining. Lastly, SEM imaging of GAS biofilms formed by this model revealed GAS cocci chains arranged into three-dimensional aggregated structures with EPS matrix material. Taken together, we outline an efficacious GAS biofilm pharyngeal cell model that can support long-term GAS biofilm formation, with biofilms formed closely resembling those seen in vivo.

scholarly journals Using 3D epigenomic maps of primary olfactory neuronal cells from living individuals to understand gene regulation

2018 ◽  
Vol 4 (12) ◽  
pp. eaav8550 ◽  
Author(s):  
Suhn K. Rhie ◽  
Shannon Schreiner ◽  
Heather Witt ◽  
Chris Armoskus ◽  
Fides D. Lay ◽  
...  
Keyword(s):  
Cell Model ◽  
Expression Profiles ◽  
Neuronal Cells ◽  
Three Dimensional ◽  
Gene Expression Profiles ◽  
Regulatory Elements ◽  
A Genome ◽  
Wide Scale ◽  
Cell Model System ◽  
Distal Regulatory Elements

As part of PsychENCODE, we developed a three-dimensional (3D) epigenomic map of primary cultured neuronal cells derived from olfactory neuroepithelium (CNON). We mapped topologically associating domains and high-resolution chromatin interactions using Hi-C and identified regulatory elements using chromatin immunoprecipitation and nucleosome positioning assays. Using epigenomic datasets from biopsies of 63 living individuals, we found that epigenetic marks at distal regulatory elements are more variable than marks at proximal regulatory elements. By integrating genotype and metadata, we identified enhancers that have different levels corresponding to differences in genetic variation, gender, smoking, and schizophrenia. Motif searches revealed that many CNON enhancers are bound by neuronal-related transcription factors. Last, we combined 3D epigenomic maps and gene expression profiles to predict enhancer-target gene interactions on a genome-wide scale. This study not only provides a framework for understanding individual epigenetic variation using a primary cell model system but also contributes valuable data resources for epigenomic studies of neuronal epithelium.

Encapsulation of Neural Cells in Nano-Featured Polymer Scaffolds through Co-axial Electrospinning

2007 ◽  
Vol 1065 ◽  
Author(s):  
Rajesh A Pareta ◽  
Thomas J Webster
Keyword(s):  
Carbon Nanotubes ◽  
Pc12 Cells ◽  
Cell Model ◽  
Electrospun Fiber ◽  
Polymer Surface ◽  
Three Dimensional ◽  
Neural Cells ◽  
Flow Rates ◽  
Tissue Organization ◽  
Controlled Flow

ABSTRACTEncapsulation of PC12 cells (neural cell model) in alginate hydrogels with a protective coating of poly(lactic-co-glycolic acid) (PLGA) was achieved in the present study using co-axial electrospinning. Co-axial electrospinning consists of two concentric capillaries compared to only one capillary in conventional electrospinning. This allows for the processing of two liquid solutions simultaneously. Neural cells suspended in hydrogels were injected in the inner capillary, while the carbon nanotubes (added for conductivity) suspended in PLGA were injected in the outer capillary at controlled flow rates. On the application of a high voltage, a compound jet formed at the capillary exits and resulted in a co-electrospun fiber of nerve cells encapsulated in PLGA with carbon nanotubes. Carbon nanotubes were included to make the outer shell conductive to stimulate the PC12 cells. In this study, the voltage varied from 0 to 15 kV and various flow rates were tested to achieve a stable cone-jet mode in electrospinning. The cell density in the media varied from 0.5 to5 million cells/ml and the polymer solution (PLGA) concentration varied from 1 to 10 mg/ml. This resulted in a three dimensional conductive scaffold with nano-features (due to carbon nanotubes) on the polymer surface, which were collected on the grounded substrate. PC12 cells were found to be viable inside microspheres after 3 days. The size of the microspheres was quite uniform and less than 200 μm. This technique may be very useful for the development of cell encapsulated scaffolds which mimic natural body tissue organization for tissue engineering applications such as nervous system regeneration.

scholarly journals Pleiotropic effects of FGFR1 on cell proliferation, survival, and migration in a 3D mammary epithelial cell model

2005 ◽  
Vol 171 (4) ◽  
pp. 663-673 ◽  
Author(s):  
Wa Xian ◽  
Kathryn L. Schwertfeger ◽  
Tracy Vargo-Gogola ◽  
Jeffrey M. Rosen
Keyword(s):  
Cell Proliferation ◽  
Epithelial Cell ◽  
Mammary Epithelial Cell ◽  
Molecular Mechanisms ◽  
Cell Model ◽  
Smooth Muscle Actin ◽  
Three Dimensional ◽  
Mammary Epithelial ◽  
Matrix Metalloproteinase 3

Members of the fibroblast growth factor (FGF) family and the FGF receptors (FGFRs) have been implicated in mediating various aspects of mammary gland development and transformation. To elucidate the molecular mechanisms of FGFR1 action in a context that mimics polarized epithelial cells, we have developed an in vitro three-dimensional HC11 mouse mammary epithelial cell culture model expressing a drug-inducible FGFR1 (iFGFR1). Using this conditional model, iFGFR1 activation in these growth-arrested and polarized mammary acini initially led to reinitiation of cell proliferation, increased survival of luminal cells, and loss of cell polarity, resulting in the disruption of acinar structures characterized by the absence of an empty lumen. iFGFR1 activation also resulted in a gain of invasive properties and the induction of matrix metalloproteinase 3 (MMP-3), causing the cleavage of E-cadherin and increased expression of smooth muscle actin and vimentin. The addition of a pan MMP inhibitor abolished these phenotypes but did not prevent the effects of iFGFR1 on cell proliferation or survival.

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