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Quantifying Adhesive Interactions Between Cells and Extracellular Matrix by Single-cell Force Spectroscopy

Quantifying Adhesive Interactions Between Cells and Extracellular Matrix by Single-cell Force Spectroscopy
Author:
Publisher:
Total Pages:
Release: 2009
Genre:
ISBN:

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Interactions of cells with their environment regulate important cellular functions and are required for the organization of cells into tissues and complex organisms. These interactions involve different types of adhesion receptors. Interactions with extracellular matrix (ECM) proteins are mainly mediated by the integrin family of adhesion molecules. Situations in which integrin-ECM interactions are deregulated cause diseases and play a crucial role in cancer cell invasion. Thus, the mechanisms underlying integrin-binding and regulation are of high interest, particularly at the molecular level. How can cell-ECM interactions be studied? While there are several methods to analyze cell adhesion, few provide quantitative data on adhesion forces. One group, single-cell force spectroscopy �ΕΠ�� ξραλϟζεζδπ αγϝδπζμλ αϟ ϟϝδ πζλϛιδ�ϐδιι ιδσδι αλγ ϐαλ ϟϝδοδεμοδ γζεεδοδλϟζαϟδ ϟϝδ αγϝδπζσδ νομνδοϟζδπ με ζλγζσζγραι ϐδιιπ� Μλδ ζκνιδκδλϟαϟζμλ με Π�ΕΠ ζπ βαπδγ μλ αϟμκζϐ εμοϐδ κζϐομπϐμνυ �ΑΕΚ�· ϟϝζπ ϟδϐϝλζξρδ ϝαπ βδδλ δκνιμυδγ ζλ ϟϝδ νοδπδλϟδγ ςμοθ� Αγσαλϟαϛδμρπιυ ΑΕΚ�Π�ΕΠ ϐμκβζλδπ ϝζϛϝ ϟδκνμοαι αλγ πναϟζαι ϐδιι καλζνριαϟζμλ� ϟϝδ αβζιζϟυ ϟμ κδαπροδ α ιαοϛδ οαλϛδ με αγϝδπζμλ εμοϐδπ αλγ πρεεζϐζδλϟιυ ϝζϛϝ�εμοϐδ οδπμιρϟζμλ ϟμ αιιμς ϟϝδ πϟργυ με πζλϛιδ�κμιδϐριδ βζλγζλϛ δσδλϟπ ζλ ϟϝδ ϐμλϟδτϟ με α ιζσζλϛ ϐδιι� Πζλϐδ ζλγζσζγραι αγϝδπζμλ οδϐδνϟμοπ ϐαλ βδ αλαιυφδγ ςζϟϝζλ ϟϝδζο νϝυπζμιμϛζϐαι δλσζομλκδλϟ� ΑΕΚ�Π�ΕΠ ζπ α νμςδοερι ϟμμι ϟμ πϟργυ ϟϝδ κδϐϝαλζπκπ ρλγδοιυζλϛ ζλϟδϛοζλ�οδϛριαϟζμλ� Ϟϝδ νοδπδλϟδγ ςμοθ ζπ πνιζϟ ζλϟμ πζτ ϐϝανϟδοπ� �ϝανϟδο μλδ ϛζσδπ βαϐθϛομρλγ ζλεμοκαϟζμλ αβμρϟ ϐδιι�Δ�Κ ζλϟδοαϐϟζμλπ� Ζλ ϐϝανϟδο ϟςμ� γζεεδοδλϟ αγϝδπζμλ αππαυπ αοδ ϐμκναοδγ αλγ ϐμλϟοαπϟδγ� Ϟϝδ ϟϝδμοδϟζϐαι Βδιι�Δσαλπ κμγδι ςϝζϐϝ ζπ ρπδγ ϟμ ζλϟδονοδϟ ζλϟδϛοζλ�κδγζαϟδγ ϐδιι αγϝδπζμλ ζπ γζπϐρππδγ ζλ ϐϝανϟδο ϟϝοδδ� Ϟϝδοδαεϟδο� ϟϝδ ϟϝοδδ νομηδϐϟπ ϟϝαϟ εμοκ ϟϝδ ϐμοδ με ϟϝδ ϟϝδπζπ αοδ γδϟαζιδγ ζλ ϐϝανϟδοπ εμρο ϟϝομρϛϝ πζτ� Ζλ ϟϝδ εζοπϟ νομηδϐϟ �ϐϝανϟδο ʹ�� �ł“�ø»�ζλϟδϛοζλ κδγζαϟδγ ϐδιι αγϝδπζμλ ϟμ ϐμιιαϛδλ ϟυνδ Ζ� ϟϝδ κμπ�


Quantifying Cell-Ecm Adhesion by Single Cell Force Spectroscopy

Quantifying Cell-Ecm Adhesion by Single Cell Force Spectroscopy
Author: Anna Taubenberger
Publisher: Sudwestdeutscher Verlag Fur Hochschulschriften AG
Total Pages: 184
Release: 2011-03
Genre:
ISBN: 9783838124278

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Integrin-mediated adhesive interactions of cells with the extracellular matrix (ECM) regulate important cellular functions and play a critical role in disease. Therefore, studying the mechanisms underlying integrin-binding and regulation is of great interest. In the presented work three respective biological questions have been addressed using Atomic Force Microscopy based Single Cell Force Spectroscopy (AFM-SCFS). The work is split into six chapters. The first chapter provides an introduction into cell-ECM interactions. Thereafter different adhesion assays are compared and contrasted, and the theoretical Bell-Evans model is discussed. In chapter four, alpha2beta1-integrin mediated cell adhesion to collagen I is quantified. Thereafter the effects of the exposure of cryptic integrin binding sites within collagen I on pre-osteoblasts are analysed. At last, the role of BCR/ABL in modulating myeloid progenitor cell adhesion to stromal cells and ECM-coated substrates is studied. Taken together, the work demonstrates that AFM-SCFS is a versatile tool that permits monitoring of cell adhesion from single-molecule recognition events to the formation of more complex adhesion sites.


Single Cell Force Spectroscopy for Quantification of Cellular Adhesion on Surfaces

Single Cell Force Spectroscopy for Quantification of Cellular Adhesion on Surfaces
Author: Wayne B Christenson
Publisher:
Total Pages: 223
Release: 2016
Genre: Cell adhesion
ISBN:

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Cell adhesion is an important aspect of many biological processes. The atomic force microscope (AFM) has made it possible to quantify the forces involved in cellular adhesion using a technique called single cell force spectroscopy (SCFS). AFM based SCFS offers versatile control over experimental conditions for probing directly the interaction between specific cell types and specific proteins, surfaces, or other cells. Transmembrane integrins are the primary proteins involved in cellular adhesion to the extra cellular matix (ECM). One of the chief integrins involved in the adhesion of leukocyte cells is alphaMbeta2 (Mac-1). The experiments in this dissertation quantify the adhesion of Mac-1 expressing human embryonic kidney (HEK Mac-1), platelets, and neutrophils cells on substrates with different concentrations of fibrinogen and on fibrin gels and multi-layered fibrinogen coated fibrin gels. It was shown that multi-layered fibrinogen reduces the adhesion force of these cells considerably. A novel method was developed as part of this research combining total internal reflection microscopy (TIRFM) with SCFS allowing for optical microscopy of HEK Mac-1 cells interacting with bovine serum albumin (BSA) coated glass after interacting with multi-layered fibrinogen. HEK Mac-1 cells are able to remove fibrinogen molecules from the multi-layered fibrinogen matrix. An analysis methodology for quantifying the kinetic parameters of integrin-ligand interactions from SCFS experiments is proposed, and the kinetic parameters of the Mac-1 fibrinogen bond are quantified. Additional SCFS experiments quantify the adhesion of macrophages and HEK Mac-1 cells on functionalized glass surfaces and normal glass surfaces. Both cell types show highest adhesion on a novel functionalized glass surface that was prepared to induce macrophage fusion. These experiments demonstrate the versatility of AFM based SCFS, and how it can be applied to address many questions in cellular biology offering quantitative insights.


Adhesive Interactions in Normal and Transformed Cells

Adhesive Interactions in Normal and Transformed Cells
Author: Yury A. Rovensky
Publisher: Springer Science & Business Media
Total Pages: 241
Release: 2011-08-31
Genre: Science
ISBN: 1617793043

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Adhesive Interactions in Normal and Transformed Cells describes the basic mechanisms of the ability of tissue cells to attach to each other and to the extracellular matrix. These adhesive interactions are pivotal regulators of main cellular functions, such as proliferation, survival and migration. The adhesive interactions are involved in embryonic development, regeneration, and also in inflammation and degeneration processes, which are at the basis of many diseases. Serious alterations in cell adhesion caused by the oncogenic transformation play a key role in cancer invasion and metastasis. This volume provides comprehensive information about structural, mechanistic and signaling aspects of adhesive interactions in both normal and cancer cells in comparison. Integration of such aspects of the adhesive process as structure, relation to cell systems of receptors and cytoskeleton, function, signaling pathways, and the alterations in tumor cells constitutes the strongest point of this work. The results of the long-time author’s research are included in the book. The author was one of pioneers, who used scanning electron microscopy (SEM) to study the cell surface morphology of normal cultured cells and the cells underwent the oncogenic transformation, processes of their attachment to and spreading on the surfaces of a solid substratum, and also surprising ability of the cells to respond to various geometric configurations of the substrata surfaces. Adhesive Interactions in Normal and Transformed Cells has both biological and medical aspects and, therefore, it can be interesting not only for cell biologists, developmental biologists and cancer researchers, but also for physicians. It is intended for researchers, postdocs, undergraduate and graduate students.


The Adhesive Interaction of Cells

The Adhesive Interaction of Cells
Author: David Garrod
Publisher: Elsevier
Total Pages: 429
Release: 1999-03-31
Genre: Science
ISBN: 0080877133

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The aim of "The Adhesive Interaction of Cells" has been to assemble a series of reviews by leading international experts embracing many of the most important recent developments in this rapidly expanding field. The purpose of all biological research is to understand the form and function of living organisms and, by comprehending the normal, to find explanations and remedies for the abnormal and for disease conditions. The molecules involved in cell adhesion are of fundamental importance to the structure and function of all multicellular organisms. In this book, the contributors focus on the systems of vertebrates, especially mammals, since these are most relevant to human disease. It would have been equally possible to concentrate on developmental processes and adhesion in lower organisms. A major function of adhesion molecules is to bind cells to each other or to the extracellular matrix, but they are much more than "glue". Adhesions in animal tissues must be dynamic-forming, persisting, or declining in regulated fashion- to facilitate the mobility and turnover of tissue cells. Moreover, the majority of adhesion molecules are transmembrane molecules and thus provide links between the cells and their surroundings. This gives rise to another major function of adhesion molecules, the capacity to transduce signals across the hydrophobic barrier imposed by the plasma membrane. Such signal transduction is crucially important to many aspects of cellular function including the regulation of cell motility, gene expression, and differentiation. The work in this book progresses through four sections. Part I discusses the four major families of adhesion molecules themselves, the integrins (Green and Humphries), the cadherins (Stappert and Kemler), the selectins (Tedder et al.) and the immunoglobulin superfamily (Simmons); part 2 considers junctional complexes involved in cell interactions: focal adhesions and adherens junctions (Ben Ze'ev), desmosomes (Garrod et al.), and tight junctions (Citi and Cordenonsi). The signaling role of adhesion molecules is the focus of part 3, through integrins and the extracellular matrix (Edwards and Streuli), through platelet adhesion (Du and Ginsberg), and in the nervous system (Hemperley). In part 4, the aim is to show how adhesive phenomena contribute to important aspects of cell behavior and human health. Leukocyte trafficking (Haskard et al.), cancer metastasis (Marshall and Hart), cell migration (Paleck et al.), and implantation and placentation (Damsky et al.) are the topics considered in depth. The different sections are, of course, not mutually exclusive: it is both undesirable and impossible to separate structure from function when considering cell adhesion. Each chapter has its unique features, but some overlap is both invevitable and valuable since it provides different perspectives on closely related topics. We hope that the whole contributes a valuable and stimulating consideration of this important topic.


Physics of Cancer

Physics of Cancer
Author: Claudia Mierke
Publisher: Iph001
Total Pages: 500
Release: 2018-10-24
Genre: Science
ISBN: 9780750317511

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This revised second edition is improved linguistically with multiple increases of the number of figures and the inclusion of several novel chapters such as actin filaments during matrix invasion, microtubuli during migration and matrix invasion, nuclear deformability during migration and matrix invasion, and the active role of the tumor stroma in regulating cell invasion.


Methods in Cell-Matrix Adhesion

Methods in Cell-Matrix Adhesion
Author: Josephine Adams
Publisher: Academic Press
Total Pages: 461
Release: 2002-06-12
Genre: Science
ISBN: 0080524966

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Critically acclaimed for more than 25 years, the Methods in Cell Biology series provides an indispensable tool for the researcher. Each volume is carefully edited by experts to contain state-of-the-art reviews and step-by-step protocols. Techniques are described completely so that methods are made accessible to users. This volume, Methods of Cell-Matrix Adhesion, contains integrated coverage on cell-matrix adhesion methods. It brings the classical methodologies and the latest techniques together in one concise volume. This coverage includes experimental protocols and their conceptual background for all aspects of cell-matrix adhesion research: the extracellular matrix, adhesion receptors, and the growing number of functional applications of matrix-adhesion in molecular cell biology. Also covered is the purification of the extracellular matrix to functional analyses of cellular responses.


Surface and Interfacial Aspects of Cell Adhesion

Surface and Interfacial Aspects of Cell Adhesion
Author: Alain Carre
Publisher: CRC Press
Total Pages: 546
Release: 2011-01-19
Genre: Science
ISBN: 9004190805

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Cell adhesion comes into play in almost all domains of life. The range of situations in which it occurs, involving organisms, living tissues, microorganisms or single cells, is endless. Cell adhesion is involved in the binding of a cell to a surface, extracellular matrix, or another cell using cell adhesion molecules. It is crucial in the formation


Cellular Analysis by Atomic Force Microscopy

Cellular Analysis by Atomic Force Microscopy
Author: Malgorzata Lekka
Publisher: CRC Press
Total Pages: 217
Release: 2017-07-06
Genre: Medical
ISBN: 1315341158

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Despite substantial evidence showing the feasibility of Atomic Force Microscopy (AFM) to identify cells with altered elastic and adhesive properties, the use of this technique as a complementary diagnostic method remains controversial. This book is designed to be a practical textbook that teaches how to assess the mechanical characteristics of living, individual cells by AFM. Following a step-by-step approach, it introduces the methodology of measurements in the case of both determination of elastic properties and quantification of adhesive properties.