Alterations in the temporal expression and function of cadherin-7 inhibit cell migration and condensation during chondrogenesis of chick limb mesenchymal cells in vitro

2009 ◽  
Vol 221 (1) ◽  
pp. 161-170 ◽  
Author(s):  
Dongkyun Kim ◽  
Shin-Sung Kang ◽  
Eun-Jung Jin
Keyword(s):  
Cell Migration ◽  
Mesenchymal Cells ◽  
Temporal Expression ◽  
And Function

Zyxin Regulates Cell Migration and Differentiation in EMT during Chicken AV Valve Morphogenesis

2013 ◽  
Vol 19 (4) ◽  
pp. 842-854 ◽  
Author(s):  
Na Li ◽  
Richard L. Goodwin ◽  
Jay D. Potts
Keyword(s):  
Cell Migration ◽  
Collagen Gel ◽  
Mesenchymal Cells ◽  
Valve Development ◽  
Focal Adhesion Protein ◽  
Endocardial Cell ◽  
Mesenchymal Transformation ◽  
And Function ◽  
Valve Formation

AbstractDuring heart valve development, epithelial–mesenchymal transformation (EMT) is a key process for valve formation. EMT leads to the generation of mesenchymal cells that will eventually become the interstitial cells (fibroblasts) of the mature valve. During EMT, cell architecture and motility change markedly; significant changes are also observed in various signaling pathways. Here we systematically examined the expression, localization, and function of zyxin, a focal adhesion protein, in EMT during atrioventricular (AV) valve morphogenesis. Expression and localization studies showed that zyxin was expressed in the AV canal region during crucial stages of valve development. An in vitro 3D collagen gel culture system was used to determine zyxin function either after siRNA gene knockdown or after overexpression. Our studies revealed that zyxin overexpression inhibited endocardial cell migration and cell differentiation and also led to a decrease in the number of migrating mesenchymal cells. Moreover, correlative cytoskeletal changes were apparent in response to both overexpression and knockdown treatments. Thus, zyxin appears to play a role as a regulator of cell migration and differentiation during EMT in chicken AV valve formation.

scholarly journals Virus-Induced Cell Motility

1998 ◽  
Vol 72 (2) ◽  
pp. 1235-1243 ◽  
Author(s):  
Christopher M. Sanderson ◽  
Michael Way ◽  
Geoffrey L. Smith
Keyword(s):  
Cell Migration ◽  
Cell Motility ◽  
Uv Light ◽  
Cell Metabolism ◽  
Cell Movement ◽  
Time Lapse ◽  
Late Gene Expression ◽  
Infected Cells ◽  
And Function

ABSTRACT Many viruses induce profound changes in cell metabolism and function. Here we show that vaccinia virus induces two distinct forms of cell movement. Virus-induced cell migration was demonstrated by an in vitro wound healing assay in which infected cells migrated independently into the wound area while uninfected cells remained relatively static. Time-lapse microscopy showed that the maximal rate of migration occurred between 9 and 12 h postinfection. Virus-induced cell migration was inhibited by preinactivation of viral particles with trioxsalen and UV light or by the addition of cycloheximide but not by addition of cytosine arabinoside or rifampin. The expression of early viral genes is therefore necessary and sufficient to induce cell migration. Following migration, infected cells developed projections up to 160 μm in length which had growth-cone-like structures and were frequently branched. Time-lapse video microscopy showed that these projections were formed by extension and condensation of lamellipodia from the cell body. Formation of extensions was dependent on late gene expression but not the production of intracellular enveloped (IEV) particles. The requirements for virus-induced cell migration and for the formation of extensions therefore differ from each other and are distinct from the polymerization of actin tails on IEV particles. These data show that poxviruses encode genes which control different aspects of cell motility and thus represent a useful model system to study and dissect cell movement.

Abstract 315: Surgical Pen Dye Inhibits Cell Migration From Saphenous Vein Explants

2014 ◽  
Vol 34 (suppl_1) ◽  
Author(s):  
Shinsuke Kikuchi ◽  
Richard D Kenagy ◽  
Alexander W Clowes
Keyword(s):  
Cell Migration ◽  
Crystal Violet ◽  
Primary Response ◽  
Alternative Methods ◽  
Vein Grafts ◽  
Significant Difference ◽  
Vein Bypass ◽  
Fetal Bovine ◽  
And Function

Markers containing dyes such as crystal violet (CAS 548-62-9) are routinely used on the adventitia of vein bypass grafts to avoid twisting during placement. Since little is known about how these dyes affect vein graft healing and function, we determined the effect of crystal violet on cell migration, a primary response to injury after grafting. Human saphenous veins were split into adventitia and media after removal of endothelium. Cell migration from 2.5 square mm explants in DMEM with 20% fetal bovine serum was measured daily for 8 days as either 1) % migration positive explants (≥ 1 cell/explant), which only measures migration, or 2) the number of cells/explant, which measures a combination of migration and proliferation. Dye was extracted from explants with ethanol and quantified by spectrophotometry. Cell migration by both methods was significantly less from blue, compared to non-blue, adventitial explants (P<.05, mean±SEM, N=11 veins with 15-30 explants/condition; Figures A and B). No medial explants were visibly blue, and there was no significant difference in migration of cells from medial explants of blue and non-blue sections of vein (data not shown). Blue adventitial explants had 65.9±8.0 ng dye/explant compared to 2.1±1.3 for non-blue explants (mean±SEM, N=7-11 veins). Dye applied in vitro to either adventitial or medial explants dose-dependently inhibited migration (IC50=21.5 ng/explant). Conclusion: Crystal violet is toxic and inhibits venous adventitial cell migration; alternative methods should be considered for marking vein grafts.

scholarly journals Patterning of Endothelial Cells and Mesenchymal Stem Cells by Laser-Assisted Bioprinting to Study Cell Migration

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Jean-Michel Bourget ◽  
Olivia Kérourédan ◽  
Manuela Medina ◽  
Murielle Rémy ◽  
Noélie Brunehilde Thébaud ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Migration ◽  
Network Architecture ◽  
Mesenchymal Cell ◽  
Mesenchymal Cells ◽  
Specific Pattern ◽  
Trophic Factors ◽  
Collagen Hydrogel ◽  
Interesting Approach

Tissue engineering of large organs is currently limited by the lack of potent vascularizationin vitro. Tissue-engineered bone grafts can be prevascularizedin vitrousing endothelial cells (ECs). The microvascular network architecture could be controlled by printing ECs following a specific pattern. Using laser-assisted bioprinting, we investigated the effect of distance between printed cell islets and the influence of coprinted mesenchymal cells on migration. When printed alone, ECs spread out evenly on the collagen hydrogel, regardless of the distance between cell islets. However, when printed in coculture with mesenchymal cells by laser-assisted bioprinting, they remained in the printed area. Therefore, the presence of mesenchymal cell is mandatory in order to create a pattern that will be conserved over time. This work describes an interesting approach to study cell migration that could be reproduced to study the effect of trophic factors.

scholarly journals The Role of MCPIP1 in Ischemia/Reperfusion Injury-Induced HUVEC Migration and Apoptosis

2015 ◽  
Vol 37 (2) ◽  
pp. 577-591 ◽  
Author(s):  
Tiebing Zhu ◽  
Qi Yao ◽  
Xiaonan Hu ◽  
Chen Chen ◽  
Honghong Yao ◽  
...  
Keyword(s):  
Cell Migration ◽  
Ischemia Reperfusion Injury ◽  
Umbilical Vein ◽  
Ischemia Reperfusion ◽  
Mitogen Activated Protein Kinase ◽  
Mitogen Activated Protein ◽  
Myocardial Ischemia Reperfusion ◽  
Umbilical Vein Endothelial Cells ◽  
And Function

Background/Aims: Monocyte chemotactic protein-induced protein 1 (MCPIP1) plays a crucial role in various cellular processes, including neurogenesis. However, the relationship between MCPIP1 and myocardial ischemia/reperfusion (I/R) injury remained illdefined. In this study, we explored whether the I/R-mediated up-regulation of MCPIP1 is critical in the modulation of both cell migration and apoptosis in human umbilical vein endothelial cells (HUVECs). Methods: Using Western blot analysis and quantitative real-time PCR, the protein expression and mRNA transcription, respectively, of MCPIP1 was detected in HUVECs. To investigate cell migration, an in vitro scratch assay and a nested matrix model were applied. Results: I/R increased the expression of MCPIP1 via the activation of the mitogen-activated protein kinase (MAPK) and PI3K/Akt pathways. I/R increased migration and apoptosis of HUVECs, which were significantly inhibited by MCPIP1 siRNA. Conclusion: These findings suggest that I/R-mediated up-regulation of MCPIP1 regulates migration and apoptosis in HUVECs. Understanding the regulation of MCPIP1 expression and function may aid in the development of an adjunct therapeutic strategy in the treatment of individuals with I/R injury.

Human Mesenchymal Cells Control T Regulatory Phenotypes and Functions.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 2308-2308
Author(s):  
Mauro Di Ianni ◽  
Beatrice Del Papa ◽  
Lorenzo Moretti ◽  
Maria De Ioanni ◽  
Elisabetta Bonifacio ◽  
...  
Keyword(s):  
T Cell ◽  
Suppressor Cells ◽  
In Vitro Study ◽  
Foxp3 Expression ◽  
Mesenchymal Cells ◽  
Regulatory Function ◽  
Cell Populations ◽  
And Function ◽  
Cd127 Expression

Abstract Despite much investigation into T regulatory cells (Tregs), little is known about the mechanism controlling their recruitment and function. Since mesenchymal cells (MSCs) exert an immune regulatory function and suppress T cell proliferation, this in vitro study investigated their role in Treg recruitment and function. hMSCs and different T cell populations (CD3+, CD3+/CD45RA+, CD3+/CD45RO+, CD4+/CD25+, CD4+/CD25+/CD45RO+, CD4+/CD25+/CD45RA+) from healthy donors were co-cultured for up to 15 days. Harvested lymphocytes were analysed by flow-cytometry; FoxP3 and CD127 expression were measured by real time PCR; regulatory activity was assessed. Within CD3+ fraction, percentages of CD4+/CD25bright and CD4+/CD25bright/Foxp3+ cells were higher in the presence of hMSCs (42% vs 34% for the CD4+/CD25bright and 20.5 vs 3.5 for the CD4+/CD25bright/Foxp3+). Within CD3+/CD45RA+ and CD3+/CD45RO+ fractions, the T reg starting fraction of the naïve population rose from 0.05% ± 0.01 CD4/CD25 positive cells to 0.2% ± 0.14 and the T reg starting fraction of the memory population rose from 0.3% ± 0.05 CD4/CD25 positive cells to 1.5% ± 0.9. Within CD4+/CD25+, CD4+/CD25+/CD45RA+, CD4+/CD25+/CD45RO+, FoxP3 expression did not change in CD4+/CD25+ cells. It increased 5.38±2.3 fold in CD4+/CD25+/CD45RA+ and 7.98±1.9 fold in CD4+/CD25+/CD45RO+ cells. CD127 expression decreased by -582±29.2 fold in CD4+/CD25+ cells, by -216±17.5 fold in CD4+/CD25+/CD45RA+ and by -71.95±12.6 fold in CD4+/CD25+/CD45RO+ cells. Cytofluorimetric analysis showed CD4+/CD25+/FoxP3+ was up-regulated to 14%±4 and CD127 down-regulated to 4.4%±1.5 vs respectively 0.2%±0.28 and 21.9%±3.5 in controls without MSCs (CD4+/CD25+ alone). FoxP3 up-regulation was more marked in CD4+/CD25bright cells (51%±10 vs 0.1%±0.7 in controls). Sorted Tregs exert potent immunosuppressive activity on CD4+/CD25- cell populations. Inhibitory activity was lost within 15 days when sorted T regs were cultured without hMSC. On day +15 proliferation using CD4+/CD25+ cells as suppressor cells was 1.7±0.8 vs 45.2±12 in controls; proliferation using CD4/CD25+/CD45RA+ as suppressor cells was 5.4±3.1, vs 21.3±11.2 in controls, and proliferation with CD4+/CD25+/RO+ as suppressor cells was 2.3±2 vs 33.5±8.7 in controls. We demonstrate MSC recruit Tregs from a fraction of CD3+ and from immunoselected CD3+/CD45RA+ and CD3+/CD45RO+ fractions. After culture with MSCs both immunoselected fractions registered increases in the CD4+/CD25bright/FoxP3 subset and CD127 expression was down regulated. When purified Treg populations (CD4/CD25+, CD4/CD25+/CD45RA+ and CD4/CD25+/CD45RO+) were used as fraction for hMSC co-cultures, they maintain FoxP3 expression and CD127 expression is down-regulated. Treg suppressive capacity was maintained in Treg populations that were layered on MSC for up to 15 days while control Tregs lost all suppressive activity after 5 days culture. In conclusion, our study demonstrates that MSCs recruit, regulate and maintain T regulatory phenotype and function over time.

CO-Culture with Mesenchymal Cells Modulates TGF-Beta/Smad And Mapk Pathways in T Regulatory Cells

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 676-676 ◽  
Author(s):  
Mauro Di Ianni ◽  
Beatrice Del Papa ◽  
Lorenzo Moretti ◽  
Simonetta Piattoni ◽  
Debora Cecchini ◽  
...  
Keyword(s):  
T Regulatory Cells ◽  
Central Element ◽  
Mesenchymal Cells ◽  
Negative Regulator ◽  
Mitogen Activated Protein Kinase ◽  
Regulatory Cells ◽  
Tgf Beta ◽  
Mapk Pathways ◽  
And Function

Abstract We previously demonstrated that when human bone marrow-derived mesenchymal cells (hMSCs) were co-cultured with CD4+/CD25+ T regulatory cells (Tregs) they maintained the T regulatory phenotype and function over time. Here we studied the TGF-beta/Smad signalling and mitogen-activated protein kinase (MAPK) pathways after Tregs/hMSCs co-culture. After 7 days co-culture of highly purified, immuno-selected Tregs and hMSCs from healthy donors, TGF-beta and IL-10 concentrations were dosed and mRNA of a panel of genes (NFAT, Smad4, Smad7, AP-1, Foxp3 and CD127) was quantified on harvested lymphocytes. Western blotting was performed by incubation with antiSMAD2, anti-pSMAD2, anti-ERK1/2 and anti-pERK1/2 polyclonal antibodies. After immuno-magnetic cell separation the final Treg fraction showed a mean purity of 93.6%±1. The CD4+/CD25+bright constituted 29%±7 of Tregs, FoxP3 cells constituted 51.9%±15.1 and CD127+ cells 19%±11.5. CD4+/CD25+ cells inhibited CD4+/CD25− cells (mean inhibition percentage: 52.1±29.6% (ratio 1:1). Tregs produced no TGF-beta and only a small quantity of IL-10 (8.67±4 pg/ml). hMSCs produced high quantity of TGF-beta (229.3 ± 54.8 pg/ml) associated with little IL-10 (2.7 ± 1.2 pg/ml). After co-culture, the TGF-beta concentration was 91.5±48.5 pg/ml while the IL-10 concentration was no different to baseline. To identify TGF-beta signaling pathways, we focused on Smad2, a central element in the cascade. In Tregs the strongly expressed pSmad2 signal after selection rapidly decreased after 7 days culture. When Tregs were co-cultured in presence of hMSCs the phosphorylated form of Smad2 did not change, indicating TGF-beta signaling was up-regulated. After co-culture mRNA quantification showed hMSCs down-regulated expression of SMAD7 (−8.9± 4.4 times vs Tregs without hMSCs), a negative regulator of TGF-beta signaling. After co-culture with hMSCs non-regulatory CD4+/CD25− control cells did not show any differences in SMAD7 expression. To study the MAPK kinase pathways, harvested Tregs were assayed for ERK1/2 kinase activity by determining their phosphorylated forms. pERK1/2 was almost absent in selected Tregs; it rapidly increased after 7 days in vitro culture without hMSCs but remained low after co-culture with hMSCs, indicating impaired ERK1/2 activation. AP-1 activation was also reduced (AP-1 mRNA −32±29 times less than controls). In conclusion, co-cultures of hMSCs and Tregs have the potential to set up a positive feedback loop that heightens responsiveness to TFG-beta in Treg cells. In fact, hMSCs produce TGF-beta which is consumed in co-culture with Tregs; TGF-beta1 signaling in Tregs is maintained through pSMAD2 up-regulation and SMAD7 down-regulation; hMSCs also inhibit ERK phosphorylation which consequently down-regulates AP-1 mRNA. Thus Tregs signalling is maintained through culture on a layer of hMSCs. Manipulation of Tregs signaling through culture on a layer of hMSCs may represent a novel strategy for maintenance of Treg phenotype and function.

Neutrophil migration through in vitro interstitial matrix

1992 ◽  
Vol 50 (1) ◽  
pp. 894-895
Author(s):  
J. Roemer ◽  
S.R. Simon
Keyword(s):  
Extracellular Matrix ◽  
Smooth Muscle ◽  
Cell Migration ◽  
Smooth Muscle Cells ◽  
Inflammatory Cell ◽  
Neutrophil Migration ◽  
Culture Conditions ◽  
Aortic Smooth Muscle ◽  
Specific Culture

We are developing an in vitro interstitial extracellular matrix (ECM) system for study of inflammatory cell migration. Falcon brand Cyclopore membrane inserts of various pore sizes are used as a support substrate for production of ECM by R22 rat aortic smooth muscle cells. Under specific culture conditions these cells produce a highly insoluble matrix consisting of typical interstitial ECM components, i.e.: types I and III collagen, elastin, proteoglycans and fibronectin.

Molecular architecture and functions of the neuronal cytoskeleton

1990 ◽  
Vol 48 (3) ◽  
pp. 2-3
Author(s):  
Nobutaka Hirokawa
Keyword(s):  
Major Elements ◽  
Molecular Structures ◽  
Organelle Transport ◽  
Molecular Architecture ◽  
Neuronal Morphogenesis ◽  
Microtubule Associated Proteins ◽  
Associated Proteins ◽  
And Function ◽  
Small Clusters

In this symposium I will present our studies about the molecular architecture and function of the cytomatrix of the nerve cells. The nerve cell is a highly polarized cell composed of highly branched dendrites, cell body, and a single long axon along the direction of the impulse propagation. Each part of the neuron takes characteristic shapes for which the cytoskeleton provides the framework. The neuronal cytoskeletons play important roles on neuronal morphogenesis, organelle transport and the synaptic transmission. In the axon neurofilaments (NF) form dense arrays, while microtubules (MT) are arranged as small clusters among the NFs. On the other hand, MTs are distributed uniformly, whereas NFs tend to run solitarily or form small fascicles in the dendrites Quick freeze deep etch electron microscopy revealed various kinds of strands among MTs, NFs and membranous organelles (MO). These structures form major elements of the cytomatrix in the neuron. To investigate molecular nature and function of these filaments first we studied molecular structures of microtubule associated proteins (MAP1A, MAP1B, MAP2, MAP2C and tau), and microtubules reconstituted from MAPs and tubulin in vitro. These MAPs were all fibrous molecules with different length and formed arm like projections from the microtubule surface.

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