Members of Prevotella Genus Distinctively Modulate Innate Immune and Barrier Functions in a Human Three-Dimensional Endometrial Epithelial Cell Model

Abstract Background Prevotella species are commonly isolated from the reproductive tract of women with obstetric/gynecologic health complications. However, contributions of this genus to changes in local microenvironment are not well characterized. Our objective was to evaluate species-specific effects of Prevotella on the human endometrial epithelium. Methods Thirteen Prevotella strains, originally isolated from the human oral cavity, amniotic fluid, endometrium, or vagina (including women with bacterial vaginosis), were obtained from BEI and ATCC resources. Bacteria were evaluated in silico and in vitro using human endometrial epithelial cells (EEC) grown as monolayers or a 3-dimensional (3D) model. Results Genomic characterization illustrated metabolic and phylogenetic diversity of Prevotella genus. Among tested species, P. disiens exhibited cytotoxicity. Scanning electron microscopy analysis of the 3D EEC model revealed species-specific colonization patterns and alterations of ultracellular structures. Infection with sialidase-producing P. timonensis resulted in elongated microvilli, and increased MUC3 and MUC4 expression. Infections with Prevotella species, including P. bivia, did not result in significant proinflammatory activation of EEC. Conclusions Collectively, findings indicate that Prevotella species are metabolically diverse and overall not cytotoxic or overtly inflammatory in EEC; however, these bacteria can form biofilms, alter barrier properties of the endometrial epithelium, and ultimately impact colonization of secondary colonizers.

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Organoids of the female reproductive tract

Journal of Molecular Medicine ◽

10.1007/s00109-020-02028-0 ◽

2021 ◽

Vol 99 (4) ◽

pp. 531-553 ◽

Cited By ~ 1

Author(s):

Cindrilla Chumduri ◽

Margherita Y. Turco

Keyword(s):

Reproductive Tract ◽

Personalised Medicine ◽

Three Dimensional ◽

In Vitro Models ◽

Experimental Models ◽

Female Reproductive Tract ◽

Cellular Heterogeneity ◽

Dynamic Regulation ◽

Pregnancy And Childbirth

AbstractHealthy functioning of the female reproductive tract (FRT) depends on balanced and dynamic regulation by hormones during the menstrual cycle, pregnancy and childbirth. The mucosal epithelial lining of different regions of the FRT—ovaries, fallopian tubes, uterus, cervix and vagina—facilitates the selective transport of gametes and successful transfer of the zygote to the uterus where it implants and pregnancy takes place. It also prevents pathogen entry. Recent developments in three-dimensional (3D) organoid systems from the FRT now provide crucial experimental models that recapitulate the cellular heterogeneity and physiological, anatomical and functional properties of the organ in vitro. In this review, we summarise the state of the art on organoids generated from different regions of the FRT. We discuss the potential applications of these powerful in vitro models to study normal physiology, fertility, infections, diseases, drug discovery and personalised medicine.

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An optimised GAS-pharyngeal cell biofilm model

Scientific Reports ◽

10.1038/s41598-021-87377-7 ◽

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.

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Interaction between vascular endothelial cells and vascular intimal spindle-shaped cells in vitro

Journal of Cell Science ◽

10.1242/jcs.60.1.89 ◽

1983 ◽

Vol 60 (1) ◽

pp. 89-102

Author(s):

D de Bono ◽

C. Green

Keyword(s):

Extracellular Matrix ◽

Endothelial Cells ◽

Vascular Endothelial Cells ◽

Three Dimensional ◽

Vascular Endothelial ◽

Pure Cultures ◽

Species Specific ◽

Endothelial Growth Factor

The interactions between human or bovine vascular endothelial cells and fibroblast-like vascular intimal spindle-shaped cells have been studied in vitro, using species-specific antibodies to identify the different components in mixed cultures. Pure cultures of endothelial cells grow as uniform, nonoverlapping monolayers, but this growth pattern is lost after the addition of spindle cells, probably because the extracellular matrix secreted by the latter causes the endothelial cells to modify the way they are attached to the substrate. The result is a network of tubular aggregates of endothelial cells in a three-dimensional ‘polylayer’ of spindle-shaped cells. On the other hand, endothelial cells added to growth-inhibited cultures of spindle-shaped cells will grow in sheets over the surface of the culture. Human endothelial cells grown in contact with spindle-shaped cells have a reduced requirement for a brain-derived endothelial growth factor. The interactions of endothelial cells and other connective tissue cells in vitro may be relevant to the mechanisms of endothelial growth and blood vessel formation in vivo, and emphasize the potential importance of extracellular matrix in controlling endothelial cell behaviour.

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

The Journal of Cell Biology ◽

10.1083/jcb.200505098 ◽

2005 ◽

Vol 171 (4) ◽

pp. 663-673 ◽

Cited By ~ 89

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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Three-Dimensional Spheroid Cell Model of In Vitro Adipocyte Inflammation

Tissue Engineering Part A ◽

10.1089/ten.tea.2014.0531 ◽

2015 ◽

Vol 21 (11-12) ◽

pp. 1837-1847 ◽

Cited By ~ 23

Author(s):

Paul A. Turner ◽

Yi Tang ◽

Stephen J. Weiss ◽

Amol V. Janorkar

Keyword(s):

Cell Model ◽

Three Dimensional

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In vitro 3D Model of Female Reproductive Tract: An Overview and Future Aspect

10.20944/preprints202108.0562.v1 ◽

2021 ◽

Author(s):

Arnaud Martino Capuzzo

Keyword(s):

Reproductive Tract ◽

Three Dimensional ◽

Reproductive Organs ◽

Experimental Models ◽

Female Reproductive Tract ◽

Complex Interactions ◽

Current State ◽

Pregnancy And Delivery ◽

Recent Developments

Hormones must be balanced and dynamically controlled for the Female Reproductive Tract (FRT) to function correctly during the menstrual cycle, pregnancy, and delivery. Gamete selection and successful transfer to the uterus, where it implants and pregnancy occurs, is supported by the mucosal epithelial lining of the FRT ovaries, uterus, cervix, fallopian tubes, and vagina. Successful implantation and placentation in humans and other animals rely on complex interactions between the embryo and a receptive female reproductive system. The FRT's recent breakthroughs in three-dimensional (3D) organoid systems now provide critical experimental models that match the organ's physiological, functional, and anatomical characteristics in vitro. This article summarizes the current state of the art on organoids generated from various parts of the FRT. The current analysis examines recent developments in the creation of organoid models of reproductive organs, as well as their future directions.

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Histopathological features of SARS-CoV-2 infection and relationships with organoid technology

Journal of International Medical Research ◽

10.1177/03000605211044382 ◽

2021 ◽

Vol 49 (9) ◽

pp. 030006052110443

Author(s):

İrem İnanç ◽

Esra Erdemli

Keyword(s):

Viral Entry ◽

Three Dimensional ◽

Human Cells ◽

Host Cells ◽

Angiotensin Converting Enzyme 2 ◽

Antiviral Compounds ◽

Global Pandemic ◽

Species Specific

Coronavirus disease 2019 (COVID-19) following infection by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has caused a global pandemic that is still having serious effects worldwide. This virus, which targets the lungs in particular, can also damage other tissues. Angiotensin converting enzyme 2 (ACE-2) plays a key role in viral entry into host cells. The presence of ACE-2 in various tissues may permit viral infection. Studies of COVID-19 often make use of postmortem tissues. Although this information provides various useful results, it is also necessary to conduct in vitro studies to understand optimal treatment approaches. Because the virus may show species-specific differences, in vitro technologies using human cells are particularly important. Organoid technologies, three-dimensional structures that can be obtained from human cells, are playing increasingly important roles in studies of SARS-CoV-2. This technology offers a significant advantage in terms of mimicking in vivo tissue structures and testing antiviral compounds. In this mini-review, we summarize studies of SARS-CoV-2 using both histopathological and organoid technology approaches.

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Self-Organogenesis from 2D Micropatterns to 3D Biomimetic Biliary Trees

10.20944/preprints202106.0579.v1 ◽

2021 ◽

Author(s):

Emilie Gontran ◽

Lorena Loarca ◽

Cyrille El Khassis ◽

Latifa Bouzhir ◽

Dmitry Ayollo ◽

...

Keyword(s):

Drug Testing ◽

Cell Attachment ◽

Cell Model ◽

Three Dimensional ◽

Culture Conditions ◽

Biliary System ◽

Biliary Duct ◽

Glass Slides ◽

Normal Rat

Background & Aims: Globally, liver diseases account for 2 million deaths per year. For those with advanced liver disease the only curative approach is liver transplantation. However, less than 10% of those in need get a liver transplant due to limited organ availability. To circumvent this challenge, there has been a great focus in generating a bioengineered liver. Despite its essential role in liver functions, a functional biliary system has not yet been developed. In this framework, exploration of epithelial cell self-organogenesis and microengineering-driven geometrical cell confinement allow to envision the bioengineering of a functional biomimetic intrahepatic biliary tract. Approach: Three-dimensional (3D) bile ducts were built in vitro by restricting cell adhesion to two-dimensional (2D) patterns to guide cell self-organization. Tree shapes mimicking the configuration of the human biliary system were micropatterned on glass slides, restricting cell attachment to these areas. Different tree geometries and culture conditions were explored to stimulate self-organogenesis of normal rat cholangiocytes (NRCs) used as a biliary cell model, either alone or in coculture with human umbilical endothelial cells (HUVECs). Results: Pre-seeding the micropatterns with HUVECs promoted luminogenesis with higher efficiency to yield functional branched biliary tubes. Lumen formation, apico-basal polarity, and preservation of the cholangiocyte phenotype were confirmed. Moreover, intact and functional biliary structures were detached from the micropatterns for further manipulation. Conclusion: This study presents physiologically relevant 3D biliary duct networks built in vitro from 2D micropatterns. This opens opportunities for investigating bile duct organogenesis, physiopathology, and drug testing.

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Generation of rat lungs by blastocyst complementation in Fgfr2b-deficient mouse model

10.1101/2022.01.05.475149 ◽

2022 ◽

Author(s):

Shunsuke Yuri ◽

Yuki Murase ◽

Aayako Isotani

Keyword(s):

Mouse Model ◽

Three Dimensional ◽

Cell Types ◽

Lung Epithelial Cells ◽

Developmental Timing ◽

Deficient Mouse ◽

Blastocyst Complementation ◽

Multiple Cell ◽

Species Specific

Regenerative medicine is a tool to compensate for the shortage of lungs for transplantation, but it remains difficult to construct a lung in vitro due to the complex three-dimensional structures and multiple cell types required. A blastocyst complementation method using interspecies chimeric animals has been attracting attention as a way to create complex organs in animals, but successful lung formation has not yet been achieved. Here, we applied a reverse-blastocyst complementation method to clarify the conditions required to form lungs in an Fgfr2b-deficient mouse model. We then successfully formed a rat-derived lung in the mouse model without generating a mouse line by applying a tetraploid-based organ-complementation method. Importantly, rat lung epithelial cells retained their developmental timing even in the mouse body. This result provides useful insights regarding the need to overcome the barrier of species-specific developmental timing in order to generate functional lungs in interspecies chimeras.

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