On-chip Neural differentiation of PC12 cells

SIEVEWELL™ 20 μm was coated with Cellmatrix® Type IV (Collagen Type IV, Nitta Gelatin Inc). Rat pheochromocytoma cell line PC12 cells were loadetd into SIEVEWELL™ 20 μm. Cells were cultured with RPMI 1640/10%horse serum/5% fetal bovine serum containing 10 ng/μL NGF. After 7 days, cells were fixed with PFA, permiabilized with 0.05% Tween 20/PBS, blocked wtih 1%BSA/PBS, stained wtih mouse anti-rat Tubulin β3 (TUBB3) antibody (Clone, TUJ1) followed by staining with Alexa Fluor Plus 488 labelled anti mouse IgG antibody and DAPI. Images were taken with THUNDER Imaging Systems (Leica Microsystems).