scholarly journals Comparison of the collagenous products synthesized in culture by pig aortic endothelial and smooth-muscle cells. Variability in endothelial-cell cultures

1984 ◽  
Vol 218 (1) ◽  
pp. 11-18 ◽  
Author(s):  
E A Sankey ◽  
M J Barnes
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Muscle Cells ◽  
Collagen Type I ◽  
Type I ◽  
Type Iv ◽  
Endothelial Cell Cultures ◽  
Membrane Type ◽  
Type V

In contrast with smooth-muscle cells from the same tissue, endothelial cells from pig aorta were found to exhibit in culture considerable variability in the pattern of collagen synthesis between one isolation of cells and the next. Synthesis varied from largely collagen type I to virtually all type III in the absence of type I but with small amounts still of collagens types IV and V, to, in one instance, synthesis basically of only type V. Synthesis usually by these cells of collagen predominantly of the interstitial type (I and III) rather than, as might be expected, that from basement membrane (type IV) was not attributable to the influence of subculture. All four collagen types were deposited in the cell layer to an increased extent in primary compared with secondary cultures of either smooth muscle or endothelial origin. Endothelial cells appeared sometimes to synthesize a large-Mr collagenous entity that might conceivably be related to ‘short-chain’ collagen. In addition, small-Mr hydroxyproline-containing peptides were detected that might reflect rapid collagen(s) turnover in endothelial cultures.

Abstract 418: Co-Culture of Endothelial Cells, Smooth Muscle Cells and Monocytes in Collagen Scaffold: A Novel i n vitro Model of Atherosclerosis

2017 ◽  
Vol 37 (suppl_1) ◽  
Author(s):  
Martin Liu ◽  
Angelos Karagiannis ◽  
Matthew Sis ◽  
Srivatsan Kidambi ◽  
Yiannis Chatzizisis
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Type I Collagen ◽  
Smooth Muscle Actin ◽  
Muscle Cells ◽  
Type I ◽  
Collagen Gels ◽  
Human Coronary Artery

Objectives: To develop and validate a 3D in-vitro model of atherosclerosis that enables direct interaction between various cell types and/or extracellular matrix. Methods and Results: Type I collagen (0.75 mg/mL) was mixed with human artery smooth muscle cells (SMCs; 6x10 5 cells/mL), medium, and water. Human coronary artery endothelial cells (HCAECs; 10 5 /cm 2 ) were plated on top of the collagen gels and activated with oxidized low density lipoprotein cholesterol (LDL-C). Monocytes (THP-1 cells; 10 5 /cm 2 ) were then added on top of the HCAECs. Immunofluorescence showed the expression of VE-cadherin by HCAECs (A, B) and α-smooth muscle actin by SMCs (A). Green-labelled LDL-C particles were accumulated in the subendothelial space, as well as in the cytoplasm of HCAECs and SMCs (C). Activated monocytes were attached to HCAECs and found in the subendothelial area (G-I). Both HCAECs and SMCs released IL-1β, IL-6, IL-8, PDGF-BB, TGF-ß1, and VEGF. Scanning and transmission electron microscopy showed the HCAECs monolayer forming gap junctions and the SMCs (D-F) and transmigrating monocytes within the collagen matrix (G-I). Conclusions: In this work, we presented a novel, easily reproducible and functional in-vitro experimental model of atherosclerosis that has the potential to enable in-vitro sophisticated molecular and drug development studies.

scholarly journals Collagen degradation and platelet-derived growth factor stimulate the migration of vascular smooth muscle cells

2000 ◽  
Vol 113 (11) ◽  
pp. 2055-2064
Author(s):  
E. Stringa ◽  
V. Knauper ◽  
G. Murphy ◽  
J. Gavrilovic
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Smooth Muscle Cell ◽  
Vascular Smooth Muscle Cells ◽  
Muscle Cell ◽  
Collagen Type ◽  
Muscle Cells ◽  
Collagen Type I ◽  
Type I

Cell migration is a key event in many biological processes and depends on signals from both extracellular matrix and soluble motogenic factors. During atherosclerotic plaque development, vascular smooth muscle cells migrate from the tunica media to the intima through a basement membrane and interstitial collagenous matrix and proliferate to form a neointima. Matrix metalloproteinases have previously been implicated in neointimal formation and in this study smooth muscle cell adhesion and migration on degraded collagen have been evaluated. Vascular smooth muscle cells adhered to native intact collagen type I and to its first degradation by-product, 3/4 fragment (generated by collagenase-3 cleavage), unwound at 35 degrees C to mimic physiological conditions. PDGF-BB pre-treatment induced a fourfold stimulation of smooth muscle cell motility on the collagen 3/4 fragment whereas no increase in smooth muscle cell motility on collagen type I was observed. Cell migration on collagen type I was mediated by alpha2 integrin, whereas PDGF-BB-stimulated migration on the 3/4 collagen fragment was dependent on alphavbeta3 integrin. alphavbeta3 integrin was organised in clusters concentrated at the leading and trailing edges of the cells and was only expressed when cells were exposed to the 3/4 collagen fragment. Tyrphostin A9, an inhibitor of PDGF receptor-beta tyrosine kinase activity, resulted in complete abolition of migration of PDGF-BB treated cells on collagen type I and 3/4 fragment. These results strongly support the hypothesis that the cellular migratory response to soluble motogens can be regulated by proteolytic modification of the extracellular matrix.

scholarly journals IL-8 reduces VCAM-1 secretion of smooth muscle cells by increasing p-ERK expression when 3-D co-cultured with vascular endothelial cells

2011 ◽  
Vol 34 (3) ◽  
pp. 138 ◽  
Author(s):  
Zhi Zhang ◽  
Guang Chu ◽  
Hong-Xian Wu ◽  
Ni Zou ◽  
Bao-Gui Sun ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Type I Collagen ◽  
Vascular Endothelial Cells ◽  
Umbilical Vein ◽  
Muscle Cells ◽  
Type I ◽  
Vascular Endothelial ◽  
Culture Model

Objective: The goal of this study was to investigate the crosstalk between vascular endothelial cells (ECs) and smooth muscle cells (SMCs) using a three-dimensional (3-D) co-culture model. In addition, the role of IL-8 in this crosstalk was investigated. Methods: A 3-D co-culture model was constructed using a Transwell chamber system and type I collagen gel. Human umbilical artery smooth muscle cells (HUASMCs) were suspended in the gel and added to the upper compartment of the Transwell. Human umbilical vein endothelial cells (HUVECs) were then grown on the surface of the gel. The growth of HUASMCs was tested with a CFDA SE cell proliferation kit. IL-8 and other bioactive substances were investigated by ELISA and real-time PCR. The alteration of p-ERK expression related to the change in IL-8 levels was also examined by Western blot analysis. Results: The proliferation rate of HUASMCs in the 3-D co-culture model was 0.679 ± 0.057. Secretion and transcription of VEGF, t-PA, NO and VCAM-1 in the 3-D co-culture model were different than in single (2-D) culture. When 3-D co-cultured, IL-8 released by HUVECs was significantly increased (2.35 ± 0.16 fold) (P﹤0.05) and the expression of VCAM-1 from HUASMCs was reduced accordingly (0.55±0.09 fold). In addition, increasing or decreasing the level of IL-8 changed the level of p-ERK and VCAM-1 expression. The reduction of VCAM-1, resulting from increased IL-8, could be blocked by the MEK inhibitor, PD98059. Conclusion: Crosstalk between HUVECs and HUASMCs occurred and was probably mediated by IL-8 in this 3-D co-culture model.

In Vitro Growth of Human Urinary Tract Smooth Muscle Cells on Laminin and Collagen Type I-Coated Membranes under Static and Dynamic Conditions

2005 ◽  
Vol 11 (1-2) ◽  
pp. 161-171 ◽  
Author(s):  
Ulrich Hubschmid ◽  
Phaik-Mooi Leong-Morgenthaler ◽  
Aurelia Basset-Dardare ◽  
Sylvie Ruault ◽  
Peter Frey
Keyword(s):  
Smooth Muscle ◽  
Urinary Tract ◽  
Smooth Muscle Cells ◽  
Collagen Type ◽  
Muscle Cells ◽  
Collagen Type I ◽  
Type I ◽  
In Vitro Growth ◽  
Coated Membranes

SIRT1 deacetylates RFX5 and antagonizes repression of collagen type I (COL1A2) transcription in smooth muscle cells

2012 ◽  
Vol 428 (2) ◽  
pp. 264-270 ◽  
Author(s):  
Jun Xia ◽  
Xiaoyan Wu ◽  
Yuyu Yang ◽  
Yuhao Zhao ◽  
Mingming Fang ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Collagen Type ◽  
Muscle Cells ◽  
Collagen Type I ◽  
Type I

Imbalance in the Synthesis of Collagen Type I and Collagen Type III in Smooth Muscle Cells Derived from Human Varicose Veins

2001 ◽  
Vol 38 (6) ◽  
pp. 560-568 ◽  
Author(s):  
Patricia Sansilvestri-Morel ◽  
Alain Rupin ◽  
Cécile Badier-Commander ◽  
Patrick Kern ◽  
Jean-Noël Fabiani ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Collagen Type ◽  
Varicose Veins ◽  
Muscle Cells ◽  
Collagen Type I ◽  
Type I ◽  
Collagen Type Iii ◽  
Type Iii

Identification Of Scavenger Receptor Sr-Bi In The Endothelial Cells And The Smooth Muscle Cells Of Rat Aorta In Vitro

1999 ◽  
Vol 5 (S2) ◽  
pp. 1338-1339
Author(s):  
Y. C. Yeh ◽  
G. Y. Hwang ◽  
V. C. Yang
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Scavenger Receptor ◽  
Muscle Cells ◽  
Rat Aorta ◽  
Cell Surface Receptor ◽  
Complete Medium ◽  
Type I ◽  
Peripheral Tissues

The class B ,type I scavenger receptor (SR-BI) was the first molecularly well defined cell surface receptor which binds HDL and mediates the selective uptake of HDL cholesteryl ester. It is expressed primarily in liver and steroidogenic tissues. However, the recent studies also suggested a potentially important role of SR-BI in the initial steps of cholesterol efflux in the peripheral tissues. In this study, we have used immunoblotting and immunofluorescence microscopy to study the expression and subcellular localization of SR-BI in the cultured endothelial cells and the smooth muscle cells of rat aorta.A peptide containing residues 495 to 509 from mSR-BI plus an NH2-terminal cysteine was coupled to hemocyanin. Then the peptide was used to generate mSR-BI495 antiserum in New Zealand white rabbits. The cells from the aortic ring of 1-month-old Spraque-Dawly rats were grown in the dishes containing complete medium (Dulbecco's modified Eagle's medium with 10% fetal bovine serum) and incubated at 37°C in 95% air/ 10 % CO2 atmosphere for 3-4 days.

Identification of Scavenger Receptor Sr-Bi in The Endothelial Cells and the Smooth Muscle Cells of Rat Aorta in Vitro and in Vivo

2000 ◽  
Vol 6 (S2) ◽  
pp. 484-485
Author(s):  
Y P. Liu ◽  
Y. C. Yeh ◽  
V. C. Yang
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Scavenger Receptor ◽  
Muscle Cells ◽  
Rat Aorta ◽  
Cell Surface Receptor ◽  
Type I

The class B, type I scavenger receptor (SR-BI) was the first molecularly well defined cell surface receptor which binds HDL and mediates the selective uptake of HDL cholesteryl ester. It is expressed primarily in liver and steroidogenic tissues. However, the recent studies also suggested a potentially important role of SR-BI in the initial steps of cholesterol efflux in the peripheral tissues. In this study, we have used immunofluorescence and immunoelectron microscopy to study the expression and subcellular localization of SR-BI in the endothelial cells and the smooth muscle cells of rat aorta in vitro and in vivo.

scholarly journals An In situ Collagen-HA Hydrogel System Promotes Survival and Preserves the Proangiogenic Secretion of hiPSC-derived Vascular Smooth Muscle Cells

2020 ◽  
Author(s):  
Biraja C. Dash ◽  
Kaiti Duan ◽  
Hao Xing ◽  
Themis R. Kyriakides ◽  
Henry C. Hsia
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Smooth Muscle Cells ◽  
Collagen Type ◽  
Muscle Cells ◽  
Collagen Type I ◽  
Type I ◽  
Hydrogel System

AbstractHuman induced pluripotent stem cell-derived vascular smooth muscle cells (hiPSC-VSMCs) with proangiogenic properties have huge therapeutic potential. While hiPSC-VSMCs have already been utilized for wound healing using a biomimetic collagen scaffold, an in situ forming hydrogel mimicking the native environment of skin offers the promise of hiPSC-VSMC mediated repair and regeneration. Herein, the impact of a collagen type-I-hyaluronic acid (HA) in situ hydrogel cross-linked using a PEG-based cross-linker on hiPSC-VSMCs viability and proangiogenic paracrine secretion was investigated. Our study demonstrated increases in cell viability, maintenance of phenotype and proangiogenic growth factor secretion, and proangiogenic activity in response to the conditioned medium. The optimally cross-linked and functionalized collagen type-I/HA hydrogel system developed in this study shows promise as an in situ hiPSC-VSMC carrier system for wound regeneration.

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