Egm 2

Pericytes were seeded at 8000 cells/mL into two 6‐well cell culture plate (Corning Inc., Corning, NY, USA) precoated with gelatin (Cell Biologics Inc.) in either Dulbecco's modified Eagle media (DMEM) with 4.5 g·L⁻¹ glucose, l‐glutamine, and sodium pyruvate (Corning Inc.) supplemented with +5% FBS (Corning Inc.) +1% penicillin/streptomycin (P/S) (Corning Inc.) or endothelial growth medium 2 (EGM‐2; Cell Biologics, Inc., Chicago, IL, USA) supplemented with the ECs medium supplement kit (Catalog M1168, Cell Biologics Inc., Chicago, IL, USA). Cells were seeded at 10 000 cells per well into a 24‐well cell culture plate (Corning) precoated with gelatin (Cell Biologics Inc.) in cell culture media. After 6 h for cells to attach, media was changed to include LDL (Sigma) or d‐glucose (Sigma). Plates were placed into an Incucyte Zoom instrument (EssenBioscience Inc., Ann Arbor, MI, USA) integrated with the incucyte zoom 2018A software (EssenBioscience Inc., Ann Arbor, MI, USA), and growth was monitored for 5 days to construct proliferation curves.

Passage 7 mouse coronary artery ECs (Cell Biologics, Inc.) were grown on Transwell filters of 0.4‐µm pores with 6.5 mm diameter (Corning Inc.) at 1 000 000 cells/mL in 200 µL per well of EGM‐2 (Cell Biologics, Inc.). PCs were grown on the bottom of the Transwell filter adaptor (Applied Biophysics Inc.) coated with gelatin (Applied Biophysics Inc.) at 500 000 cells/mL in 1 mL of DMEM + 5% FBS + 1% P/S. Inserts were placed into a Transwell filter adapter and then moved to the ECIS Ztheta 16W array module (Applied Biophysics Inc., Troy, NY, USA) inside a cell incubator at 37 °C and 5% CO₂. TEER measured for 48 h. After 48 h, media was changed and TEER was measured for another 48 h. Analysis was done by the ECIS software. Data were normalized to the cell‐free chamber.