How long should the cells attach before starting the flow?

The cells must be firmly attached to the bottom of the channel slide before the flow can be started. Adherence time is dependent on the cell type, medium supplements, and surface. When using HUVEC on a collagen-coated surface, for example, the adherence time is only half an hour. After this, shear stress can be applied. In order to give the cells the opportunity to adapt to the new environmental conditions, a stepwise increase of the shear stress is recommended. Details for HUVEC are given in Application Note 13 "Endothelial Cells Under Perfusion" (PDF).

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