Linear Measurement of Cell Contraction in a Capillary Collagen Gel System

Ilagan, Roger; Guthrie, Kelly; Quinlan, Sarah; Rapoport, H. Scott; Jones, Sarah; Church, Ashley; Basu, Joydeep; Ludlow, John W.

doi:10.2144/000113349

Cited by 17 publications

(17 citation statements)

References 12 publications

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“…The collagen gel assay was performed following a published procedure 33 . A hydrogel solution was prepared in a serum-free environment with the following: 10X MEM (cat# 11430, Invitrogen), sodium bicarbonate (cat# S5761, Sigma-Aldrich), L-glutamine (cat# G6392, Sigma-Aldrich) and HEPES buffer (cat# 15630, Invitrogen).…”

Section: Methodsmentioning

confidence: 99%

Interaction of cochlin and mechanosensitive channel TREK-1 in trabecular meshwork cells influences the regulation of intraocular pressure

Carreon

Castellanos

Gasull

et al. 2017

Sci Rep

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In the eye, intraocular pressure (IOP) is tightly regulated and its persistent increase leads to ocular hypertension and glaucoma. We have previously shown that trabecular meshwork (TM) cells might detect aqueous humor fluid shear stress via interaction of the extracellular matrix (ECM) protein cochlin with the cell surface bound and stretch-activated channel TREK-1. We provide evidence here that interaction between both proteins are involved in IOP regulation. Silencing of TREK-1 in mice prevents the previously demonstrated cochlin-overexpression mediated increase in IOP. Biochemical and electrophysiological experiments demonstrate that high shear stress-induced multimeric cochlin produces a qualitatively different interaction with TREK-1 compared to monomeric cochlin. Physiological concentrations of multimeric but not monomeric cochlin reduce TREK-1 current. Results presented here indicate that the interaction of TREK-1 and cochlin play an important role for maintaining IOP homeostasis.

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Section: Methodsmentioning

confidence: 99%

Interaction of cochlin and mechanosensitive channel TREK-1 in trabecular meshwork cells influences the regulation of intraocular pressure

Carreon

Castellanos

Gasull

et al. 2017

Sci Rep

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“…SMCs spontaneously induce contraction of a collagen matrix in a Ca 2 + -dependant manner when embedded in a three-dimensional gel. 22,23 As shown in Figure 4, representative samples of adipose-and peripheral blood-derived cells contract to a degree comparable to bladder-derived SMCs and this contractility is inhibited by EDTA, a known Ca 2 + chelator.…”

Section: Resultsmentioning

confidence: 96%

“…Contraction percentage was normalized using the + EDTA negative controls as a background subtracted value to account for Ca 2 + -independent contraction of the collagen gels. 23 …”

Section: Contractile Assaymentioning

confidence: 99%

Regeneration of Native-Like Neo-Urinary Tissue from Nonbladder Cell Sources

Basu

Jayo

Ilagan

et al. 2012

Tissue Engineering Part A

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Urinary pathology requiring urinary diversion, partial or full bladder replacement, is a significant clinical problem affecting ~14,000 individuals annually in the United States alone. The use of gastrointestinal tissue for urinary diversion or bladder reconstruction/replacement surgeries is frequently associated with complications. To try and alleviate or reduce the frequency of these complications, tissue engineering and regenerative medicine strategies have been developed using bio-absorbable materials seeded with cells derived from the bladder. However, bladder-sourced cells may not always be suitable for such applications, especially in patients with bladder cancer. In this study, we describe the isolation and characterization of smooth muscle cells (SMCs) from porcine adipose and peripheral blood that are phenotypically and functionally indistinguishable from bladder-derived SMCs. In a preclinical Good Laboratory Practice study, we demonstrate that autologous adipose- and peripheral blood-derived SMCs may be used to seed synthetic, biodegradable tubular scaffold structures and that implantation of these seeded scaffolds into a porcine cystectomy model leads to successful de novo regeneration of a tubular neo-organ composed of urinary-like neo-tissue that is histologically identical to native bladder. The ability to create urologic structures de novo from scaffolds seeded by autologous adipose- or peripheral blood-derived SMCs will greatly facilitate the translation of urologic tissue engineering technologies into clinical practice.

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“…Others have reduced fibroblast contraction of collagen gels through the chelation of divalent cations (e.g. Ca 2 + ) (Ilagan et al, 2010) or the antagonism of endogenous TGFβ signalling or heparin sulfate-containing proteoglycan synthesis (Chen et al, 2005). However, such interventions would have direct effects on the recruitment process itself, and thus be inappropriate to use in models of lymphocyte migration.…”

Section: Discussionmentioning

confidence: 99%

Analysis of the effects of stromal cells on the migration of lymphocytes into and through inflamed tissue using 3-D culture models

Jeffery

Buckley

Moss

et al. 2013

Journal of Immunological Methods

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Stromal cells may regulate the recruitment and behaviour of leukocytes during an inflammatory response, potentially through interaction with the endothelial cells (EC) and the leukocytes themselves. Here we describe new in vitro methodologies to characterise the effects of stromal cells on the migration of lymphocytes through endothelium and its underlying matrix. Three-dimensional tissue-like constructs were created in which EC were cultured above a stromal layer incorporating fibroblasts either as a monolayer on a porous filter or dispersed within a matrix of collagen type 1. A major advantage of these constructs is that they enable each step in leukocyte migration to be analysed in sequence (migration through EC and then stroma), as would occur in vivo. Migrated cells can also be retrieved from the constructs to identify which subsets traffic more effectively and how their functional responses evolve during migration. We found that culture of EC with dermal fibroblasts promoted lymphocyte transendothelial migration but not onward transit through matrix. A critical factor influencing the effect of fibroblasts on recruitment proved to be their proximity to the EC, with direct contact tending to disrupt migration. Comparison of the different approaches indicates that choice of an appropriate 3-D model enables the steps in lymphocyte entry into tissue to be studied in sequence, the regulatory mechanism to be dissected, and the effects of changes in stroma to be investigated.

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Linear Measurement of Cell Contraction in a Capillary Collagen Gel System

Cited by 17 publications

References 12 publications

Interaction of cochlin and mechanosensitive channel TREK-1 in trabecular meshwork cells influences the regulation of intraocular pressure

Interaction of cochlin and mechanosensitive channel TREK-1 in trabecular meshwork cells influences the regulation of intraocular pressure

Regeneration of Native-Like Neo-Urinary Tissue from Nonbladder Cell Sources

Analysis of the effects of stromal cells on the migration of lymphocytes into and through inflamed tissue using 3-D culture models

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