Rotating wall vessel as a new in vitro shear stress generation system: application to rat coronary endothelial cell cultures.
In this paper, we describe a simple new design for the application of controlled, top-hat profiled wall shear stress forces in a way that is independent of hydrostatic pressure and oxygen tension, based on a rotating wall vessel system. This system has been applied to the culture of rat coronary endothelial cells obtained with a Langendorff-derived procedure isolation. Endothelial cells are immunopurified on the basis of RECA expression, and conservation of endothelial phenotype has been assessed on the basis of morphology, RECA and von Willebrand factor expressions and diI-Ac-LDL uptake. Shear stress induced by the rotating wall vessel was measured using a mathematical formula specifically designed for this type of model, and its impact on coronary endothelial cells was evaluated. Shear stress produced cell orientation parallel to the flux direction, elevated NO production and decreased monocyte adhesion. Cells were kept viable and functional for at least 4 days under shear. This simple design allows the handling and management of numerous vials in parallel and appears to be suitable for large-scale studies of both the acute and chronic impact of modulation of the physico-chemical environment on endothelial cell physiology and function.
Morin
J P
JP
INSERM E9920, Faculté de Médecine Pharmacie, IFRMP 23, Rouen, France. jp.morin@univ-rouen.fr
Preterre
D
D
Keravec
V
V
Thuillez
C
C
D000818:Animals; D001808:Blood Vessels / Q000473:pathology*; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D003331:Coronary Vessels / Q000166:cytology*; D004730:Endothelium, Vascular / Q000166:cytology*; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D008077:Lipoproteins, LDL / Q000378:metabolism; D008163:Luminescent Measurements; D015233:Models, Statistical; D009000:Monocytes / Q000166:cytology / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D010641:Phenotype; D011312:Pressure; D051381:Rats; D017208:Rats, Wistar; D013314:Stress, Mechanical; D020298:U937 Cells; D014841:von Willebrand Factor / Q000096:biosynthesis
D008077:Lipoproteins, LDL; D014841:von Willebrand Factor; D009569:Nitric Oxide; D010100:Oxygen