Inhibition of urokinase synthesis and cell surface binding alters the motile behavior of embryonic endocardial-derived mesenchymal cells in vitro

Development ◽  
1993 ◽  
Vol 118 (3) ◽  
pp. 931-939 ◽  
Author(s):  
P.G. McGuire ◽  
S.M. Alexander
Keyword(s):  
Cell Surface ◽  
Mesenchymal Cells ◽  
Surface Binding ◽  
Atrioventricular Canal ◽  
Cell Surface Binding ◽  
Cell Matrix ◽  
Matrix Interactions ◽  
Mesenchymal Transformation ◽  
Cell Matrix Interactions

The expression of the serine protease urokinase is elevated during the epithelial-mesenchymal transformation of the endocardium in the developing avian heart. Elevated urokinase expression is associated with the migrating mesenchymal cells of the atrioventricular canal and bulbotruncus and not the myocardium. Treatment of isolated endocardial-derived mesenchymal cells with phosphatidyinositol-specific phospholipase C released urokinase and its receptor from the cell surface and caused significant alterations in cell morphology and motility. Likewise inhibition of urokinase synthesis by treatment of cells with antisense oligonucleotides also inhibited the migration and motility of the endocardial-derived cells. These results suggest an important role for this enzyme in cell-matrix interactions and cell migration during development.

scholarly journals N-sulfation of heparan sulfate is critical for syndecan-4-mediated podocyte cell-matrix interactions

2016 ◽  
Vol 310 (10) ◽  
pp. F1123-F1135 ◽  
Author(s):  
Terrel Sugar ◽  
Deborah J. Wassenhove-McCarthy ◽  
A. Wayne Orr ◽  
Jonette Green ◽  
Toin H. van Kuppevelt ◽  
...  
Keyword(s):  
Cell Surface ◽  
Heparan Sulfate ◽  
Focal Adhesion Kinase ◽  
Focal Adhesion ◽  
Cell Matrix ◽  
Matrix Interactions ◽  
Cell Matrix Interactions ◽  
Cell Surface Proteoglycan

Previous research has shown that podocytes unable to assemble heparan sulfate on cell surface proteoglycan core proteins have compromised cell-matrix interactions. This report further explores the role of N-sulfation of intact heparan chains in podocyte-matrix interactions. For the purposes of this study, a murine model in which the enzyme N-deacetylase/ N-sulfotransferase 1 (NDST1) was specifically deleted in podocytes and immortalized podocyte cell lines lacking NDST1 were developed and used to explore the effects of such a mutation on podocyte behavior in vitro. NDST1 is a bifunctional enzyme, ultimately responsible for N-sulfation of heparan glycosaminoglycans produced by cells. Immunostaining of glomeruli from mice whose podocytes were null for Ndst1 ( Ndst1−/−) showed a disrupted pattern of localization for the cell surface proteoglycan, syndecan-4, and for α-actinin-4 compared with controls. The pattern of immunostaining for synaptopodin and nephrin did not show as significant alterations. In vitro studies showed that Ndst1−/− podocytes attached, spread, and migrated less efficiently than Ndst1+/+ podocytes. Immunostaining in vitro for several markers for molecules involved in cell-matrix interactions showed that Ndst1−/− cells had decreased clustering of syndecan-4 and decreased recruitment of protein kinase-Cα, α-actinin-4, vinculin, and phospho-focal adhesion kinase to focal adhesions. Total intracellular phospho-focal adhesion kinase was decreased in Ndst1−/− compared with Ndst1+/+ cells. A significant decrease in the abundance of activated integrin α5β1 on the cell surface of Ndst1−/− cells compared with Ndst1+/+ cells was observed. These results serve to highlight the critical role of heparan sulfate N-sulfation in facilitating normal podocyte-matrix interactions.

scholarly journals Imaging and Analysis of Cellular Locations in Three-Dimensional Tissue Models

2019 ◽  
Vol 25 (3) ◽  
pp. 753-761 ◽  
Author(s):  
Warren Colomb ◽  
Matthew Osmond ◽  
Charles Durfee ◽  
Melissa D. Krebs ◽  
Susanta K. Sarkar
Keyword(s):  
Three Dimensional ◽  
Human Mesenchymal Stem Cells ◽  
Basic Research ◽  
Light Sheet ◽  
Cell Matrix ◽  
Matrix Interactions ◽  
Cell Matrix Interactions ◽  
Tissue Models ◽  
Alginate Matrix

AbstractThe absence of quantitative in vitro cell–extracellular matrix models represents an important bottleneck for basic research and human health. Randomness of cellular distributions provides an opportunity for the development of a quantitative in vitro model. However, quantification of the randomness of random cell distributions is still lacking. In this paper, we have imaged cellular distributions in an alginate matrix using a multiview light sheet microscope and developed quantification metrics of randomness by modeling it as a Poisson process, a process that has constant probability of occurring in space or time. We imaged fluorescently labeled human mesenchymal stem cells embedded in an alginate matrix of thickness greater than 5 mm with $\sim\! {\rm 2}{\rm. 9} \pm {\rm 0}{\rm. 4}\,\mu {\rm m}$ axial resolution, the mean full width at half maximum of the axial intensity profiles of fluorescent particles. Simulated randomness agrees well with the experiments. Quantification of distributions and validation by simulations will enable quantitative study of cell–matrix interactions in tissue models.

scholarly journals Peptide gels of fully-defined composition and mechanics for probing cell-cell and cell-matrix interactions in vitro

2020 ◽  
Vol 85-86 ◽  
pp. 15-33 ◽  
Author(s):  
J.C. Ashworth ◽  
J.L. Thompson ◽  
J.R. James ◽  
C.E. Slater ◽  
S. Pijuan-Galitó ◽  
...  
Keyword(s):  
Cell Matrix ◽  
Matrix Interactions ◽  
Cell Matrix Interactions ◽  
Cell Cell

scholarly journals An Inducible Model for Unraveling the Effects of Advanced Glycation End-Products in Collagen with Age and Diabetes

2020 ◽  
Author(s):  
Austin G. Gouldin ◽  
Jennifer L. Puetzer
Keyword(s):  
Cell Viability ◽  
Blue Light ◽  
Advanced Glycation End Products ◽  
Advanced Glycation ◽  
Cell Matrix ◽  
Matrix Interactions ◽  
Glycation End Products ◽  
End Products ◽  
Cell Matrix Interactions

AbstractIn connective tissues there is a clear link between increasing age and degeneration. It is believed advanced glycation end-products (AGEs) play a central role in this degeneration. AGEs are sugar induced non-enzymatic crosslinks which accumulate in collagen with age and diabetes, altering tissue mechanics and cellular function. Despite ample correlative evidence linking collagen glycation to degeneration, little is known how AGEs impact cell-matrix interactions, limiting therapeutic options. One reason for this limited understanding is AGEs are typically induced in vitro using high concentrations of ribose which decrease cell viability and make it impossible to investigate cell-matrix interactions. The objective of this study was to develop a system to trigger AGE accumulation while maintaining cell viability. Using cell-seeded high density collagen gels, we investigated the effect of two different systems for AGE induction, ribose at low concentrations (30, 100, and 200 mM) over 15 days of culture and riboflavin (0.25 mM and 0.75mM) induced with blue light for 40 seconds. We found ribose and riboflavin with blue light are capable of producing a wide range of AGE crosslinks which match and/or exceed reported human AGE levels for various tissues, ages, and diseases, without affecting cell viability and metabolism. Interestingly, a single 40 second treatment of riboflavin and blue light produced similar levels of AGEs as 3 days of 100 mM ribose treatment and matched aged mouse tendon AGE levels. This riboflavin treatment option is an exciting means to trigger AGE crosslinks on demand in vivo or in vitro without impacting cell metabolism or viability and holds great promise for further unraveling the mechanism of AGEs in age and diabetes related tissue degeneration.

Understanding and Regulating Cell-Matrix Interactions Using Hydrogels of Designable Mechanical Properties

2021 ◽  
Vol 17 (2) ◽  
pp. 149-168
Author(s):  
Jiapeng Yang ◽  
Yu Zhang ◽  
Meng Qin ◽  
Wei Cheng ◽  
Wei Wang ◽  
...  
Keyword(s):  
Mechanical Properties ◽  
Mechanical Response ◽  
Research Direction ◽  
Mechanical Environment ◽  
Cell Functions ◽  
Cell Matrix ◽  
Matrix Interactions ◽  
Sense And Respond ◽  
Cell Matrix Interactions

Similar to natural tissues, hydrogels contain abundant water, so they are considered as promising biomaterials for studying the influence of the mechanical properties of extracellular matrices (ECM) on various cell functions. In recent years, the growing research on cellular mechanical response has revealed that many cell functions, including cell spreading, migration, tumorigenesis and differentiation, are related to the mechanical properties of ECM. Therefore, how cells sense and respond to the extracellular mechanical environment has gained considerable attention. In these studies, hydrogels are widely used as the in vitro model system. Hydrogels of tunable stiffness, viscoelasticity, degradability, plasticity, and dynamical properties have been engineered to reveal how cells respond to specific mechanical features. In this review, we summarize recent process in this research direction and specifically focus on the influence of the mechanical properties of the ECM on cell functions, how cells sense and respond to the extracellular mechanical environment, and approaches to adjusting the stiffness of hydrogels.

Rapid generation of collagen-based microtissues to study cell–matrix interactions

TECHNOLOGY ◽  
2016 ◽  
Vol 04 (02) ◽  
pp. 80-87 ◽  
Author(s):  
Marie-Elena Brett ◽  
Alexandra L. Crampton ◽  
David K. Wood
Keyword(s):  
Large Scale ◽  
Culture Conditions ◽  
Encapsulated Cells ◽  
Cell Contractility ◽  
Tissue Constructs ◽  
Cell Matrix ◽  
Matrix Interactions ◽  
Rapid Generation ◽  
Cell Matrix Interactions

The objective of this study was to create a method for studying cell–matrix interactions in a physiologically relevant 3D protein-based tissue construct that could be scaled up to perform large-scale screens, study cell–matrix interactions on a population basis, or be remodeled by cells to build larger tissues. We have developed an easy-to-use method to miniaturize protein-based tissue constructs that maintains the 3D in vitro environment, while alleviating several obstacles associated with larger avascular tissue constructs. In this study, we demonstrate that (i) cells can interact with the 3D environment both while encapsulated or while interacting only with the surface of the microtissues, (ii) encapsulated cells are highly viable and, for the first time, (iii) microtissues on this size scale (~200 μm) can be used to quantify cell contractility. This versatile platform should facilitate large-scale screens in 3D in vitro culture conditions for drug development and high throughput mechanistic biology.

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