Fig. 1
From: A glomerulus-on-a-chip to recapitulate the human glomerular filtration barrier
Characterization of cellular lines and description of the microfluidic chip. a–i Flow cytometry for WT1 (PE, red) in hAKPC-P (a), hiPOD (b), and hpPOD (c); for nephrin (NPHS1-FITC, green) in hAKPC-P (d), hiPOD (e), and hpPOD (f) and for CD31 (FITC, green) in hAKPC-P (g), hiPOD (h), and hpPOD (i). All three lines show almost 100% expression of podocyte markers and absence of endothelial marker confirming their podocyte phenotype. j–l Flow cytometry for WT1 (PE, red; j), for nephrin (NPHS1-FITC, green; k), and for CD31 (FITC, green; l) in hGEC show low expression (<1%) of podocyte markers and higher expression of endothelial marker confirming their endothelial phenotype. m–r Flow cytometry for WT1 (PE, red) in HuLEC (m) and hFIB (p), for nephrin (NPHS1-FITC, green) in HuLEC (n) and hFIB (q), and for CD31 (FITC, green) in HuLEC (o) and hFIB (r). HuLEC and hFIB are both negative for podocyte makers, HuLEC are positive for endothelial marker while hFIB are negative. Unstained control is shown in red while stained sample is shown in light blue. s, t The chip is a microfluidic layer sandwiched between two 175 μm glass plates (OrganoPlateTM platform, courtesy of MIMETASTM, panel s). The three-channel version of the OrganoplateTM comprises 40 networks on one 96-well format plate. t The central section is subdivided into three channels by a system known as PhaseGuide™, a thin 30 μm tall ridge that acts as a pinning barrier for incoming fluids23. Following the filling of channel E with collagen I, channel C (representing the vascular space) is seeded with cells and then filled with growth medium; the channel F (representing the urinary space) is where the filtrate is collected. Cross-section depicts patterning of cells and collagen within the GOAC. Flow of culture medium is achieved by leveling between the media reservoirs of the lanes C and F. The platform is placed on an interval rocker, with an angle to assure leveling. By changing the angle of the platform (rocker settings: interval = 8 min, angle = 7°), the direction of fluid flow is reversed
