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Application of fluid shear stress

PM Paul-Lennard Mendez
LO Leon Obendorf
JJ Jerome Jatzlau
WB Wiktor Burdzinski
MR Maria Reichenbach
VN Vanasa Nageswaran
AH Arash Haghikia
VS Verena Stangl
CH Christian Hiepen
PK Petra Knaus
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Cells were seeded on gelatin-coated μ-Slide I 0.4 Luer (ibidi GmbH) at 2.5 × 106 cells/mL and cultured for 48 h to reach confluency in the full medium before the experiment with a daily medium exchange. Shear stress was applied using the ibidi pump system (ibidi GmbH) with the associated software (V. 1.5.4). Two pump systems were used to allow simultaneous application of LSS (1 dyn/cm2) and HSS (30 dyn/cm2). For long-term experiments, cells were allowed to adapt to increasing levels of shear stress in an adaption phase (2.5 h: 2.5 dyn/cm2 for 60 min, 5 dyn/cm2 for 30 min, 10 dyn/cm2 for 30 min, 20 dyn/cm2 for 30 min, 30 dyn/cm2 for 24 h). Static controls were seeded in silicone forms with the same area as μ-Slides and cultured in 10-cm dishes. For LSS and HSS, we used WHITE and YELLOW-and-GREEN perfusion sets (ibidi GmbH), respectively.

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