(A) Schematic for bioreactor. Two different syringe pumps are used, allowing independent flow rate changes to each channel. (B) CFD simulations of shear stress through 7-µm slits at a flow rate of 1.5 µL/min in each channel. The bioreactor generates uniform shear profiles across all slits, within and above the physiological shear stress range (1 - 4 dynes/cm2 in marrow sinusoids). (C) Bioreactor is positioned over a microscope equipped with real-time imaging and green fluorescence. The entire system is placed inside an incubator at 370C and 5% CO2. Media is perfused through the system before Mk loading. Image shows a cell-free system of the fabricated bioreactor. (D) Upon Mk (Calcein AM live stain) loading and shear exposure, Mks generate proPLTs through the slits that then get fragmented by the secondary shear flow (image at 20X, scale bar 50 μm).
