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Parallel-plate flow chamber and continuous flow circuit to evaluate endothelial progenitor cells under laminar flow shear stress.

Publication ,  Journal Article
Lane, WO; Jantzen, AE; Carlon, TA; Jamiolkowski, RM; Grenet, JE; Ley, MM; Haseltine, JM; Galinat, LJ; Lin, F-H; Allen, JD; Truskey, GA; Achneck, HE
Published in: Journal of visualized experiments : JoVE
January 2012

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. 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. 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. 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).

Duke Scholars

Published In

Journal of visualized experiments : JoVE

DOI

EISSN

1940-087X

ISSN

1940-087X

Publication Date

January 2012

Issue

59

Start / End Page

3349

Related Subject Headings

  • Viscosity
  • Stem Cells
  • Shear Strength
  • Humans
  • Fractionation, Field Flow
  • Endothelial Cells
  • Cytological Techniques
  • Animals
  • 3101 Biochemistry and cell biology
  • 1702 Cognitive Sciences
 

Citation

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Lane, W. O., Jantzen, A. E., Carlon, T. A., Jamiolkowski, R. M., Grenet, J. E., Ley, M. M., … Achneck, H. E. (2012). Parallel-plate flow chamber and continuous flow circuit to evaluate endothelial progenitor cells under laminar flow shear stress. Journal of Visualized Experiments : JoVE, (59), 3349. https://doi.org/10.3791/3349
Lane, Whitney O., Alexandra E. Jantzen, Tim A. Carlon, Ryan M. Jamiolkowski, Justin E. Grenet, Melissa M. Ley, Justin M. Haseltine, et al. “Parallel-plate flow chamber and continuous flow circuit to evaluate endothelial progenitor cells under laminar flow shear stress.Journal of Visualized Experiments : JoVE, no. 59 (January 2012): 3349. https://doi.org/10.3791/3349.
Lane WO, Jantzen AE, Carlon TA, Jamiolkowski RM, Grenet JE, Ley MM, et al. Parallel-plate flow chamber and continuous flow circuit to evaluate endothelial progenitor cells under laminar flow shear stress. Journal of visualized experiments : JoVE. 2012 Jan;(59):3349.
Lane, Whitney O., et al. “Parallel-plate flow chamber and continuous flow circuit to evaluate endothelial progenitor cells under laminar flow shear stress.Journal of Visualized Experiments : JoVE, no. 59, Jan. 2012, p. 3349. Epmc, doi:10.3791/3349.
Lane WO, Jantzen AE, Carlon TA, Jamiolkowski RM, Grenet JE, Ley MM, Haseltine JM, Galinat LJ, Lin F-H, Allen JD, Truskey GA, Achneck HE. Parallel-plate flow chamber and continuous flow circuit to evaluate endothelial progenitor cells under laminar flow shear stress. Journal of visualized experiments : JoVE. 2012 Jan;(59):3349.

Published In

Journal of visualized experiments : JoVE

DOI

EISSN

1940-087X

ISSN

1940-087X

Publication Date

January 2012

Issue

59

Start / End Page

3349

Related Subject Headings

  • Viscosity
  • Stem Cells
  • Shear Strength
  • Humans
  • Fractionation, Field Flow
  • Endothelial Cells
  • Cytological Techniques
  • Animals
  • 3101 Biochemistry and cell biology
  • 1702 Cognitive Sciences