Vascular Cell Basal Medium

Protocol tips

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	Vascular cell basal media (#PCS-100-030; ATCC) supplemented with 0.2% bovinebrain extract, 5 ngmL1 human epidermal growth factor,10 mM L-glutamine, 0.75 unitsmL1 heparin sulfate,1 lgmL1 hydrocortisone, 50 lgmL1 ascorbic acid, 2%fetal bovine serum and pen/strep.

Protocol tips
Vascular cell basal media (#PCS-100-030; ATCC) supplemented with 0.2% bovinebrain extract, 5 ngmL1 human epidermal growth factor,10 mM L-glutamine, 0.75 unitsmL1 heparin sulfate,1 lgmL1 hydrocortisone, 50 lgmL1 ascorbic acid, 2%fetal bovine serum and pen/strep.

Publication protocol

Primary cultures of human coronary artery endothelial cells(HCAECs; < 5 passages; #PCS-100-020; ATCC, Manassas,VA, USA) were cultured in vascular cell basal media (#PCS-100-030; ATCC) supplemented with 0.2% bovinebrain extract, 5 ngmL1 human epidermal growth factor,10 mM L-glutamine, 0.75 unitsmL1 heparin sulfate,1 lgmL1 hydrocortisone, 50 lgmL1 ascorbic acid, 2%fetal bovine serum and pen/strep. For serum starvationexperiments, HCAECs were cultured in vascular cell basalmedia (#PCS-100-030; ATCC) supplemented with 10 mML-glutamine, 0.75 unitsmL1 heparin sulfate, 1 lgmL1hydrocortisone, 50 lgmL1 ascorbic acid and pen/strep for48 h before RNA collection. For cell shape patterning, collagen I-coated coverslips and 96-well plates with micropatternssurrounded by non-adhesive surfaces (Cytoo Inc., Grenoble,France) were seeded with ~ 5000 or 50 000 HCAECs perwell and coverslip, respectively, in accordance with the manufacturer’s instructions. For the control, cells were seeded atlow density approximately equal to that seen in the micropatterned conditions (to minimize cell-to-cell contacts) oncollagen I-coated coverslips and 96-well plates. For allexperiments, disc, crossbow, H, Y, and L adhesive micropatterns (1600 lm2) plus controls were contained on the samechip or plate to reduce experimental variability.

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