Key Spec Table
|Species Reactivity||Key Applications|
|B, H, Po, Rb||ICC|
|Safety Information according to GHS|
|Storage and Shipping Information|
|Storage Conditions||When stored at 2-8º C, the kit components are stable up to the expiration date. Do not freeze or expose to elevated temperatures. Discard any remaining reagents after the expiration date.|
|Material Size||100 assays|
Actin Cytoskeleton / Focal Adhesion Staining Kit SDS
|Reference overview||Pub Med ID|
|TIMP-1 Induces an EMT-Like Phenotypic Conversion in MDCK Cells Independent of Its MMP-Inhibitory Domain.|
Young Suk Jung,Xu-Wen Liu,Rosemarie Chirco,Richard B Warner,Rafael Fridman,Hyeong-Reh Choi Kim
PloS one 7 2012
Matrix metalloproteinases (MMPs) and their endogenous inhibitors (TIMPs) regulate epithelial-mesenchymal transition (EMT) critical for the development of epithelial organs as well as cancer cell invasion. TIMP-1 is frequently overexpressed in several types of human cancers and serves as a prognostic marker. The present study investigates the roles of TIMP-1 on the EMT process and formation of the lumen-like structure in a 3D Matrigel culture of MDCK cells. We show that TIMP-1 overexpression effectively prevents cell polarization and acinar-like structure formation. TIMP-1 induces expression of the developmental EMT transcription factors such as SLUG, TWIST, ZEB1 and ZEB2, leading to downregulation of epithelial marker and upregulation of mesenchymal markers. Importantly, TIMP-1's ability to induce the EMT-like process is independent of its MMP-inhibitory domain. To our surprise, TIMP-1 induces migratory and invasive properties in MDCK cells. Here, we present a novel finding that TIMP-1 signaling upregulates MT1-MMP and MMP-2 expression, and potentiates MT1-MMP activation of pro-MMP-2, contributing to tumor cell invasion. In spite of the fact that TIMP-1, as opposed to TIMP-2, does not interact with and inhibit MT1-MMP, TIMP-1 may act as a key regulator of MT1-MMP/MMP-2 axis. Collectively, our findings suggest a model in which TIMP-1 functions as a signaling molecule and also as an endogenous inhibitor of MMPs. This concept represents a paradigm shift in the current view of TIMP-1/MT1-MMP interactions and functions during cancer development/progression.
|Effect of substrate stiffness on pulmonary fibroblast activation by TGF-β.|
H N Chia,M Vigen,A M Kasko
Acta biomaterialia 8 2012
Peptide crosslinkers containing the sequence C-X-CG (X represents various adhesive peptides) were incorporated into poly(ethylene glycol) (PEG) hydrogel networks with different mechanical properties. Pulmonary fibroblasts (PFs) exhibit increased adhesion to rigid hydrogels modified with X=RGDS, DGEA and IKVAV (0.5 and/or 5 mM) compared with a scrambled control (X=HRPNS). PFs exhibit increased adhesion to softer hydrogels when X=DGEA at low (0.5 mM) peptide concentration. PFs seeded onto hydrogels modified with X=RGDS produce alpha-smooth muscle actin (α-SMA), a myofibroblast marker, and form an extensive cytoskeleton with focal adhesions. Decreasing substrate stiffness (achieved through hydrolytic degradation) results in down-regulation of α-SMA expression by PFs. Substrate stiffness increases the sensitivity of PFs to exogenously applied transforming growth factor beta (TGF-β1); PFs on the most rigid gels (E=900 kPa) express α-SMA when treated with low concentrations of TGF-β1 (1 ng ml(-1)), while those on less rigid gels (E=20-60 kPa) do not. These results demonstrate the importance of both mechanical and chemical cues in studying pulmonary fibroblast activation, and establish PEG hydrogels as a viable material for further study of IPF etiology.
|Mitochondria-targeted peptide MTP-131 alleviates mitochondrial dysfunction and oxidative damage in human trabecular meshwork cells.|
Min Chen,Bingqian Liu,Qianying Gao,Yehong Zhuo,Jian Ge
Investigative ophthalmology & visual science 52 2011
To investigate the antioxidative ability of a novel mitochondria-targeted peptide MTP-131 in immortalized human trabecular meshwork (iHTM) and glaucomatous human trabecular meshwork (GTM(3)) cell lines.
|How to assess cytotoxicity of (iron oxide-based) nanoparticles: a technical note using cationic magnetoliposomes.|
Stefaan J H Soenen,Marcel De Cuyper
Contrast media & molecular imaging 6 2011
The range of different types of nanoparticles and their biomedical applications is rapidly growing, creating a need to thoroughly examine the effects these particles have on biological entities. One of the most commonly used nanoparticle types is iron oxide nanoparticles, which can be used as MRI contrast agents. The main research topic is the in vitro labeling of cells with iron oxide nanoparticles to render the cells detectable for MRI upon in vivo transplantation. For the correct evaluation of cell function and behavior in vivo, any effects of the nanoparticles on the cells must be completely ruled out. The present work provides a technical note where a detailed overview is given of several assays that could be useful to determine nanoparticle toxicity. The assays described focus on (i) nanoparticle internalization, (ii) immediate cell toxicity, (iii) cell proliferation, (iv) cell morphology, (v) cell functionality and (vi) cell physiology. Potential pitfalls, appropriate controls and advantages/disadvantages of the different assays are given. The main focus of this work is to provide a detailed guide to help other researchers in the field interested in setting up nanoparticle-toxicity studies.
|Balance of life and death in alveolar epithelial type II cells: proliferation, apoptosis, and the effects of cyclic stretch on wound healing.|
Lynn M Crosby,Charlean Luellen,Zhihong Zhang,Larry L Tague,Scott E Sinclair,Christopher M Waters
American journal of physiology. Lung cellular and molecular physiology 301 2011
After acute lung injury, repair of the alveolar epithelium occurs on a substrate undergoing cyclic mechanical deformation. While previous studies showed that mechanical stretch increased alveolar epithelial cell necrosis and apoptosis, the impact of cell death during repair was not determined. We examined epithelial repair during cyclic stretch (CS) in a scratch-wound model of primary rat alveolar type II (ATII) cells and found that CS altered the balance between proliferation and cell death. We measured cell migration, size, and density; intercellular gap formation; cell number, proliferation, and apoptosis; cytoskeletal organization; and focal adhesions in response to scratch wounding followed by CS for up to 24 h. Under static conditions, wounds were closed by 24 h, but repair was inhibited by CS. Wounding stimulated cell motility and proliferation, actin and vinculin redistribution, and focal adhesion formation at the wound edge, while CS impeded cell spreading, initiated apoptosis, stimulated cytoskeletal reorganization, and attenuated focal adhesion formation. CS also caused significant intercellular gap formation compared with static cells. Our results suggest that CS alters several mechanisms of epithelial repair and that an imbalance occurs between cell death and proliferation that must be overcome to restore the epithelial barrier.
|A novel shell-structure cell microcarrier (SSCM) for cell transplantation and bone regeneration medicine.|
Kai Su,Yihong Gong,Chunming Wang,Dong-An Wang
Pharmaceutical research 28 2011
The present study aims to develop a novel open and hollow shell-structure cell microcarrier (SSCM) to improve the anchorage-dependent cell (ADC) loading efficiency, increase the space for cell proliferation and tissue regeneration, and better propel its therapeutic effects.
|Manipulating location, polarity, and outgrowth length of neuron-like pheochromocytoma (PC-12) cells on patterned organic electrode arrays.|
Yu-Sheng Hsiao,Chung-Chih Lin,Hsin-Jui Hsieh,Shih-Min Tsai,Chiung-Wen Kuo,Chih-Wei Chu,Peilin Chen
Lab on a chip 11 2011
In this manuscript, we describe a biocompatible organic electrode system, comprising poly(3,4-ethylenedioxythiophene) (PEDOT) microelectrode arrays on indium tin oxide (ITO) glass, that can be used to regulate the neuron type, location, polarity, and outgrown length of neuron-like cells (PC-12). We fabricated a PEDOT microelectrode array with four different sizes (flat; 20, 50, and 100 ?m) through electrochemical polymerization. Extracellular matrix proteins absorbed well on these organic electrodes; cells absorbed selectively on the organic electrodes when we used polyethylene oxide/polypropylene oxide/polyethylene oxide triblock copolymers (PEO/PPO/PEO, Pluronic™ F108) as the anti-adhesive coating. In this system, the neurite polarities and neuron types could be manipulated by varying the width of the PEDOT microelectrode arrays. On the unpatterned PEDOT electrode, PC-12 cells were randomly polarized, with approximately 80% having multi-polar cell types. In contrast, when we cultured PC-12 cells on the 20 ?m wide PEDOT line array, the neurites aligned along the direction of the organic electrodes, with the percentage of uni- and bipolar PC-12 cells increasing to greater than 90%. The outgrowth of neurites on the microelectrodes was promoted by ~60% with an applied electrical stimulation. Therefore, these electroactive PEDOT microelectrode arrays have potential for use in tissue engineering related to the development and regeneration of mammalian nervous systems.
|In vitro cellular response and in vivo primary osteointegration of electrochemically modified titanium.|
F Ravanetti, P Borghetti, E De Angelis, R Chiesa, FM Martini, C Gabbi, A Cacchioli
Acta biomaterialia 6 1014-24 2010
Anodic spark deposition (ASD) is an attractive technique for improving the implant-bone interface that can be applied to titanium and titanium alloys. This technique produces a surface with microporous morphology and an oxide layer enriched with calcium and phosphorus. The aim of the present study was to investigate the biological response in vitro using primary human osteoblasts as a cellular model and the osteogenic primary response in vivo within a short experimental time frame (2 and 4 weeks) in an animal model (rabbit). Responses were assessed by comparing the new electrochemical biomimetic treatments to an acid-etching treatment as control. The in vitro biological response was characterized by cell morphology, adhesion, proliferation activity and cell metabolic activity. A complete assessment of osteogenic activity in vivo was achieved by estimating static and dynamic histomorphometric parameters at several time points within the considered time frame. The in vitro study showed enhanced osteoblast adhesion and higher metabolic activity for the ASD-treated surfaces during the first days after seeding compared to the control titanium. For the ASD surfaces, the histomorphometry indicated a higher mineral apposition rate within 2 weeks and a more extended bone activation within the first week after surgery, leading to more extensive bone-implant contact after 2 weeks. In conclusion, the ASD surface treatments enhanced the biological response in vitro, promoting an early osteoblast adhesion, and the osteointegrative properties in vivo, accelerating the primary osteogenic response.
|Cell culture and motility study on a polymer surface with a nanometer-scaled stripe structure.|
Sakamoto Y, Sato K, Abo M, Tsukahara T, Kitamori T, Abe K, Yoshimura E
Biosci Biotechnol Biochem 74 569-72. Epub 2010 Mar 7. 2010
We developed a large cell culture surface with a nanostripe structure by paving polydimethylsiloxane (PDMS) replicas of a glass mold. The stripe structure has a height of 180 nm and top width of 500 nm with 400-nm intervals between stripes. Human stomach cancer SH-10-TC cells cultured on the surface changed their morphology to elongated shapes parallel to the nanostripes. In addition, cell motility parallel to the stripes was greatly enhanced. These findings strongly suggest that the nanostripe structure affected the cell physiology.
|High intracellular iron oxide nanoparticle concentrations affect cellular cytoskeleton and focal adhesion kinase-mediated signaling.|
Soenen SJ, Nuytten N, De Meyer SF, De Smedt SC, De Cuyper M
Small 6 832-42. 2010
Iron oxide nanoparticle internalization exerts detrimental effects on cell physiology for a variety of particles, but little is known about the mechanism involved. The effects of high intracellular levels of four types of iron oxide particles (Resovist, Endorem, very small organic particles, and magnetoliposomes (MLs)) on the viability and physiology of murine C17.2 neural progenitor cells and human blood outgrowth endothelial cells are reported. The particles diminish cellular proliferation and affect the actin cytoskeleton and microtubule network architectures as well as focal adhesion formation and maturation. The extent of the effects correlates with the intracellular concentration (= iron mass) of the particles, with the biggest effects for Resovist and MLs at the highest concentration (1000 microg Fe mL(-1)). Similarly, the expression of focal adhesion kinase (FAK) and the amount of activated kinase (pY397-FAK) are affected. The data suggest that high levels of perinuclear localized iron oxide nanoparticles diminish the efficiency of protein expression and sterically hinder the mature actin fibers, and could have detrimental effects on cell migration and differentiation.
|Reprogramming Cell Fate and Function Novel Strategies for iPSC Generation, Characterization, and Differentiation|
|Actin Cytoskeleton and Focal Adhesion Staining Kit|
Newsletters / Publications
|Cellutions - The newsletter for Cell Biology Researchers Volume 3: 2011|