The Mechanical Influence of Densification on Initial Epithelial Architecture DOI Creative Commons
Christian Cammarota, Nicole S. Dawney,

Phillip M. Bellomio

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: May 9, 2023

Summary Epithelial tissues are the most abundant tissue type in animals, lining body cavities and generating compartment barriers. The function of a monolayer epithelium – whether protective, secretory, absorptive, or filtrative –relies on regular architecture with respect to apical-basal axis. Using an unbiased 3D analysis pipeline developed our lab, we previously showed that epithelial architectures culture can be divided into distinct developmental categories, these intimately connected cell density: at sparse densities, cultured layers have squamous morphology (Immature); intermediate develop lateral cell-cell borders rounded apices (Intermediate); cells highest densities reach their full height demonstrate flattened (Mature). These observations prompted us ask emerges from mechanical constraints densification, what extent hallmark feature cells, namely adhesion, contributes. In other words, is shape layer simple matter sticky, deformable objects squeezing together? We addressed this problem using combination computational modeling experimental manipulations. Our results show first morphological transition, Immature Intermediate, explained simply by crowding. Additionally, identify new division (and thus transition) within Intermediate category, find second relies adhesion.

Language: Английский

The mechanical influence of densification on epithelial architecture DOI Creative Commons
Christian Cammarota, Nicole S. Dawney,

Philip M. Bellomio

et al.

PLoS Computational Biology, Journal Year: 2024, Volume and Issue: 20(4), P. e1012001 - e1012001

Published: April 1, 2024

Epithelial tissues are the most abundant tissue type in animals, lining body cavities and generating compartment barriers. The function of a monolayered epithelial tissue–whether protective, secretory, absorptive, or filtrative–relies on side-by-side arrangement its component cells. mechanical parameters that determine shape cells apical-basal plane not well-understood. architecture culture is intimately connected to cell density, cultured layers transition between architectures as they proliferate. This prompted us ask what extent emerges from two considerations: A) constraints densification B) cell-cell adhesion, hallmark feature To address these questions, we developed novel polyline cell-based computational model used it make theoretical predictions about upon changes density adhesion. We tested using experiments. Our results show appearance extended lateral borders arises consequence crowding–independent However, cadherin-mediated adhesion associated with architectural transition. suggest this represents initial distinctive architecture. Together our work reveals distinct roles layer formation provides framework understand less well-studied tissues.

Language: Английский

Citations

4
Study Models for Chlamydia trachomatis Infection of the Female Reproductive Tract DOI Creative Commons
Jaehyeon Kim, Milena Ślęczkowska,

Beatriz Nobre

et al.

Microorganisms, Journal Year: 2025, Volume and Issue: 13(3), P. 553 - 553

Published: Feb. 28, 2025

Chlamydia trachomatis (Ct) is a leading cause of sexually transmitted infections globally, often resulting in inflammatory disorders, ectopic pregnancies, and infertility. Studying Ct’s pathogenesis remains challenging due to its unique life cycle host-specific interactions, which require diverse experimental models. Animal studies using mouse, guinea pig, non-human primate models provide valuable insights into immune responses, hormonal influences, disease progression. However, they face limitations terms translational relevance physiological differences, as well ethical concerns. Complementing these, vitro systems, ranging from simple monolayer advanced three-dimensional models, exhibit improved by replicating the human tissue architecture. This includes detailed investigation epithelial barrier disruptions, epithelium–stroma responses at cellular level. Nonetheless, fall short mimicking intricate structures found vivo and, therefore, cannot faithfully replicate host–pathogen interactions or infection dynamics observed living organisms. review presents comprehensive overview employed over past few decades investigate Ct pathogenesis, addressing their strengths limitations. Furthermore, we explore emerging technologies, including organ-on-chip silico promising tools overcome existing challenges refine our understanding infections.

Language: Английский

Citations

0
A novel strategy to facilitate uniform epithelial cell maturation using liquid–liquid interfaces DOI Creative Commons
Rie Sonoi, Masamichi Kamihira

Scientific Reports, Journal Year: 2024, Volume and Issue: 14(1)

Published: May 29, 2024

Epithelial tissue forms and maintains a critical barrier function in the body. A novel culture design aimed at promoting uniform maturation of epithelial cells using liquid materials is described. Culturing Madin-Darby canine kidney (MDCK) liquid-liquid interface yielded reduced migration stimulated active cell growth. Similar to solid-liquid interfaces, cultured on fibronectin-coated exhibited growth, ultimately reaching confluent state. These stress fiber formation adopted cobblestone-like shape, which led their even distribution vessel. To inhibit apoptosis, exposure interfaces Y27632, specific inhibitor Rho-associated protein kinase (ROCK), facilitated tight junction (frequency ZO-2-positive cells, FZ = 0.73). In Y27632-exposed interface, value obtained by subtracting standard deviation ratio nucleus densities each region that compartmentalized vessel from 1, denoted as HLN, was 0.93 ± 0.01, indicated t 72 h. The behavior contributes promotion maturation.

Language: Английский

Citations

1
A novel strategy to facilitate uniform epithelial cell maturation on liquid-liquid interfaces DOI Creative Commons

Rie Sonoi,

Masamichi Kamihira

Research Square (Research Square), Journal Year: 2024, Volume and Issue: unknown

Published: Feb. 8, 2024

Abstract Background: Liquid-liquid interfaces, such as lacrimal fluid of the eye, are pervasive in living organism. While culturing cells an environment that closely mimics vivo conditions enhances our understanding cell characteristics and unveils underlying mechanisms, practical considerations have led to predominant use solid surfaces, polystyrene or glass, for culture. In this study, we introduce a novel culture design aimed at promoting uniform maturation epithelial using liquid materials. Results: Culturing Madin–Darby canine kidney (MDCK) liquid-liquid interface yielded reduced migration stimulated active growth. Similar solid-liquid cultured on fibronectin-coated exhibited growth, ultimately reaching confluent state. These stress fiber formation adopted cobblestone-like shape, which their even distribution vessel. Analysis staining images tight junction (TJ) protein ZO-2 revealed frequency ZO-2-positive cells, F Z , cultures was 2.2-fold higher than non-coated t = 72 h. To inhibit apoptosis, exposure Y27632, specific inhibitor Rho-associated kinase (ROCK), facilitated ( 0.73). Additionally, similar trend observed 0.80) Y27632-exposed interface, value obtained by subtracting standard deviation ratio nucleus densities each region compartmentalized vessel from 1, denoted H LN 0.93 0.01, indicating Across all conditions, values close correlation R 2 0.93), suggesting is related TJs late phase maturation. Conclusions: Our findings suggest behaviors interfaces contribute promotion

Language: Английский

Citations

0
A novel strategy to facilitate uniform epithelial cell maturation using liquid–liquid interfaces DOI Creative Commons
Rie Sonoi, Masamichi Kamihira

Research Square (Research Square), Journal Year: 2024, Volume and Issue: unknown

Published: Feb. 28, 2024

Abstract Epithelial tissue forms and maintains a critical barrier function in the body. A novel culture design aimed at promoting uniform maturation of epithelial cells using liquid materials is described. Culturing Madin–Darby canine kidney (MDCK) liquid–liquid interface yielded reduced migration stimulated active cell growth. Similar to solid–liquid interfaces, cultured on fibronectin-coated exhibited growth, ultimately reaching confluent state. These stress fiber formation adopted cobblestone-like shape, which led their even distribution vessel. To inhibit apoptosis, exposure interfaces Y27632, specific inhibitor Rho-associated protein kinase (ROCK), facilitated tight junction (frequency ZO-2-positive cells, F Z = 0.73). In Y27632-exposed interface, value obtained by subtracting standard deviation ratio nucleus densities each region that compartmentalized vessel from 1, denoted as H LN , was 0.93 ± 0.01, indicated t 72 h. The behavior contributes promotion maturation.

Language: Английский

Citations

0
A novel strategy to facilitate uniform epithelial cell maturation using liquid–liquid interfaces DOI Creative Commons
Rie Sonoi, Masamichi Kamihira

Research Square (Research Square), Journal Year: 2024, Volume and Issue: unknown

Published: March 5, 2024

Abstract Epithelial tissue forms and maintains a critical barrier function in the body. A novel culture design aimed at promoting uniform maturation of epithelial cells using liquid materials is described. Culturing Madin–Darby canine kidney (MDCK) liquid–liquid interface yielded reduced migration stimulated active cell growth. Similar to solid–liquid interfaces, cultured on fibronectin-coated exhibited growth, ultimately reaching confluent state. These stress fiber formation adopted cobblestone-like shape, which led their even distribution vessel. To inhibit apoptosis, exposure interfaces Y27632, specific inhibitor Rho-associated protein kinase (ROCK), facilitated tight junction (frequency ZO-2-positive cells, F Z = 0.73). In Y27632-exposed interface, value obtained by subtracting standard deviation ratio nucleus densities each region that compartmentalized vessel from 1, denoted as H LN , was 0.93 0.01, indicated t 72 h. The behavior contributes promotion maturation.

Language: Английский

Citations

0
A novel strategy to facilitate uniform epithelial cell maturation using liquid–liquid interfaces DOI Creative Commons
Rie Sonoi, Masamichi Kamihira

Research Square (Research Square), Journal Year: 2024, Volume and Issue: unknown

Published: March 6, 2024

Abstract Epithelial tissue forms and maintains a critical barrier function in the body. A novel culture design aimed at promoting uniform maturation of epithelial cells using liquid materials is described. Culturing Madin–Darby canine kidney (MDCK) liquid–liquid interface yielded reduced migration stimulated active cell growth. Similar to solid–liquid interfaces, cultured on fibronectin-coated exhibited growth, ultimately reaching confluent state. These stress fiber formation adopted cobblestone-like shape, which led their even distribution vessel. To inhibit apoptosis, exposure interfaces Y27632, specific inhibitor Rho-associated protein kinase (ROCK), facilitated tight junction (frequency ZO-2-positive cells, F Z = 0.73). In Y27632-exposed interface, value obtained by subtracting standard deviation ratio nucleus densities each region that compartmentalized vessel from 1, denoted as H LN , was 0.93 0.01, indicated t 72 h. The behavior contributes promotion maturation.

Language: Английский

Citations

0
Automated Layer Analysis (ALAn): An Image Analysis Tool for the Unbiased Characterization of Mammalian Epithelial Architecture in Culture DOI Creative Commons
Christian Cammarota, Dan T. Bergstralh, Tara M. Finegan

et al.

BIO-PROTOCOL, Journal Year: 2024, Volume and Issue: 14(1343)

Published: Jan. 1, 2024

Cultured mammalian cells are a common model system for the study of epithelial biology and mechanics. Epithelia often considered as pseudo-two dimensional thus imaged analyzed with respect to apical tissue surface. We found that three-dimensional architecture monolayers can vary widely even within small culture wells, layers appear organized in plane show gross disorganization apical-basal plane. Epithelial cell shapes should be 3D understand maturity cultured accurately compare between experiments. Here, we present detailed protocol use our image analysis pipeline, Automated Layer Analysis (ALAn), developed quantitatively characterize layers. ALAn is based on set rules applied spatial distributions DNA actin signals (depth) dimension obtained from imaging using confocal microscope. facilitates reproducibility across experiments, investigations, labs, providing users quantitative, unbiased characterization maturity. Key features • This was spatially analyze an automated fashion. requires two inputs: nuclei fluorescence microscopy. code written Python3 Jupyter Notebook interactive format. Optimized Marbin-Darby Canine Kidney (MDCK) successfully human MCF-7 mammary gland-derived Caco-2 colon carcinoma cells. utilizes Imaris software segment but may adapted alternative method. centroid coordinates volume nuclei.

Language: Английский

Citations

0
The Mechanical Influence of Densification on Initial Epithelial Architecture DOI Creative Commons
Christian Cammarota, Nicole S. Dawney,

Phillip M. Bellomio

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: May 9, 2023

Summary Epithelial tissues are the most abundant tissue type in animals, lining body cavities and generating compartment barriers. The function of a monolayer epithelium – whether protective, secretory, absorptive, or filtrative –relies on regular architecture with respect to apical-basal axis. Using an unbiased 3D analysis pipeline developed our lab, we previously showed that epithelial architectures culture can be divided into distinct developmental categories, these intimately connected cell density: at sparse densities, cultured layers have squamous morphology (Immature); intermediate develop lateral cell-cell borders rounded apices (Intermediate); cells highest densities reach their full height demonstrate flattened (Mature). These observations prompted us ask emerges from mechanical constraints densification, what extent hallmark feature cells, namely adhesion, contributes. In other words, is shape layer simple matter sticky, deformable objects squeezing together? We addressed this problem using combination computational modeling experimental manipulations. Our results show first morphological transition, Immature Intermediate, explained simply by crowding. Additionally, identify new division (and thus transition) within Intermediate category, find second relies adhesion.

Language: Английский

Citations

0