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International Journal of Experimental Pathology logoLink to International Journal of Experimental Pathology
. 1992 Oct;73(5):625–632.

The development of an in-vitro perfusion system for studies on cultured cells.

V Salih 1, S E Greenwald 1, C F Chong 1, A Coumbe 1, C L Berry 1
PMCID: PMC2002021  PMID: 1419778

Abstract

The design and use of a perfusion system, using a modified flow chamber for studies on cultured animal cells, is described. Rat thoracic aorta smooth muscle cells were isolated by an explant method and grown on Thermanox coverslips. These were introduced into the flow chamber. A flow rate of 25ml/min and a shear stress of 14.6 dynes/cm2 (12 dyne = 10 microN) (both within physiological limits) were maintained. Cells remained attached to the coverslips after 8h of perfusion with culture medium. The effect of exposing rat smooth muscle cells to the cardiovascular toxin, allylamine, is also described. The components of the system are routinely available, simple to clean, easy to assemble and sterilize. The incorporation of an in-line sensor that monitors pH, PO2, PCO2 and temperature ensures that the perfusion conditions remain within physiological limits. Automation means that minimal supervision is required. This system provides a potential mechanism in which cultured vascular cells may be perfused under controlled haemodynamic conditions, and their response to a cytotoxin may be evaluated.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Chamley-Campbell J. H., Campbell G. R. What controls smooth muscle phenotype? Atherosclerosis. 1981 Nov-Dec;40(3-4):347–357. doi: 10.1016/0021-9150(81)90145-3. [DOI] [PubMed] [Google Scholar]
  2. Dewey C. F., Jr, Bussolari S. R., Gimbrone M. A., Jr, Davies P. F. The dynamic response of vascular endothelial cells to fluid shear stress. J Biomech Eng. 1981 Aug;103(3):177–185. doi: 10.1115/1.3138276. [DOI] [PubMed] [Google Scholar]
  3. Engbers G. H., Dost L., Hennink W. E., Aarts P. A., Sixma J. J., Feijen J. An in vitro study of the adhesion of blood platelets onto vascular catheters. Part I. J Biomed Mater Res. 1987 May;21(5):613–627. doi: 10.1002/jbm.820210507. [DOI] [PubMed] [Google Scholar]
  4. Grabowski E. F., Jaffe E. A., Weksler B. B. Prostacyclin production by cultured endothelial cell monolayers exposed to step increases in shear stress. J Lab Clin Med. 1985 Jan;105(1):36–43. [PubMed] [Google Scholar]
  5. Levesque M. J., Nerem R. M. The elongation and orientation of cultured endothelial cells in response to shear stress. J Biomech Eng. 1985 Nov;107(4):341–347. doi: 10.1115/1.3138567. [DOI] [PubMed] [Google Scholar]
  6. Levesque M. J., Nerem R. M. The study of rheological effects on vascular endothelial cells in culture. Biorheology. 1989;26(2):345–357. doi: 10.3233/bir-1989-26218. [DOI] [PubMed] [Google Scholar]
  7. Lieber B. B., Giddens D. P. Post-stenotic core flow behavior in pulsatile flow and its effects on wall shear stress. J Biomech. 1990;23(6):597–605. doi: 10.1016/0021-9290(90)90052-5. [DOI] [PubMed] [Google Scholar]
  8. Lundgren C. H., Herring M. B., Arnold M. P., Glover J. L., Bendick P. J. Fluid shear disruption of cultured endothelium: the effect of cell species, fibronectin cross-linking and supporting polymer. ASAIO Trans. 1986 Jul-Sep;32(1):334–338. [PubMed] [Google Scholar]
  9. Sakariassen K. S., Aarts P. A., de Groot P. G., Houdijk W. P., Sixma J. J. A perfusion chamber developed to investigate platelet interaction in flowing blood with human vessel wall cells, their extracellular matrix, and purified components. J Lab Clin Med. 1983 Oct;102(4):522–535. [PubMed] [Google Scholar]
  10. Sakariassen K. S., Bolhuis P. A., Sixma J. J. Platelet adherence to subendothelium of human arteries in pulsatile and steady flow. 1980 Aug 15-Sep 1Thromb Res. 19(4-5):547–559. doi: 10.1016/0049-3848(80)90027-4. [DOI] [PubMed] [Google Scholar]
  11. Sprague E. A., Steinbach B. L., Nerem R. M., Schwartz C. J. Influence of a laminar steady-state fluid-imposed wall shear stress on the binding, internalization, and degradation of low-density lipoproteins by cultured arterial endothelium. Circulation. 1987 Sep;76(3):648–656. doi: 10.1161/01.cir.76.3.648. [DOI] [PubMed] [Google Scholar]
  12. Wechezak A. R., Viggers R. F., Sauvage L. R. Fibronectin and F-actin redistribution in cultured endothelial cells exposed to shear stress. Lab Invest. 1985 Dec;53(6):639–647. [PubMed] [Google Scholar]
  13. Wick T. M., Moake J. L., Udden M. M., Eskin S. G., Sears D. A., McIntire L. V. Unusually large von Willebrand factor multimers increase adhesion of sickle erythrocytes to human endothelial cells under controlled flow. J Clin Invest. 1987 Sep;80(3):905–910. doi: 10.1172/JCI113151. [DOI] [PMC free article] [PubMed] [Google Scholar]

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