| Parallel-plate Flow Chamber and Continuous Flow Circuit to Evaluate Endothelial Progenitor Cells under Laminar Flow Shear Stress. | |
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MedLine Citation:
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PMID: 22297325 Owner: NLM Status: In-Data-Review |
Abstract/OtherAbstract:
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The overall goal of this method is to describe a technique to subject adherent cells to laminar flow conditions and evaluate their response to well quantifiable fluid shear stresses(1). Our flow chamber design and flow circuit (Fig. 1) contains a transparent viewing region that enables testing of cell adhesion and imaging of cell morphology immediately before flow (Fig. 11A, B), at various time points during flow (Fig. 11C), and after flow (Fig. 11D). These experiments are illustrated with human umbilical cord blood-derived endothelial progenitor cells (EPCs) and porcine EPCs(2,3). This method is also applicable to other adherent cell types, e.g. smooth muscle cells (SMCs) or fibroblasts. The chamber and all parts of the circuit are easily sterilized with steam autoclaving. In contrast to other chambers, e.g. microfluidic chambers, large numbers of cells (> 1 million depending on cell size) can be recovered after the flow experiment under sterile conditions for cell culture or other experiments, e.g. DNA or RNA extraction, or immunohistochemistry (Fig. 11E), or scanning electron microscopy(5). The shear stress can be adjusted by varying the flow rate of the perfusate, the fluid viscosity, or the channel height and width. The latter can reduce fluid volume or cell needs while ensuring that one-dimensional flow is maintained. It is not necessary to measure chamber height between experiments, since the chamber height does not depend on the use of gaskets, which greatly increases the ease of multiple experiments. Furthermore, the circuit design easily enables the collection of perfusate samples for analysis and/or quantification of metabolites secreted by cells under fluid shear stress exposure, e.g. nitric oxide (Fig. 12)(6). |
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Authors:
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Whitney O Lane; Alexandra E Jantzen; Tim A Carlon; Ryan M Jamiolkowski; Justin E Grenet; Melissa M Ley; Justin M Haseltine; Lauren J Galinat; Fu-Hsiung Lin; Jason D Allen; George A Truskey; Hardean E Achneck |
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Publication Detail:
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Type: Journal Article Date: 2012-01-17 |
Journal Detail:
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Title: Journal of visualized experiments : JoVE Volume: - ISSN: 1940-087X ISO Abbreviation: J Vis Exp Publication Date: 2012 |
Date Detail:
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Created Date: 2012-02-02 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101313252 Medline TA: J Vis Exp Country: United States |
Other Details:
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Languages: eng Pagination: - Citation Subset: IM |
Affiliation:
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Department of Surgery, Duke University Medical Center. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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