MCDB 131 Medium

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	Culture the HMEC-1 (ATCC CRL-3243) cells at 37 °C in a 7% CO2 humidified atmosphere using MCDB-131 medium with phenol red and supplemented with 15% fetal calf serum, penicillin, streptomycin, L-glutamine 10 mM, hydrocortisone 1 μg/mL, endothelial cell growth supplement (20 μg/mL).

Protocol tips
Culture the HMEC-1 (ATCC CRL-3243) cells at 37 °C in a 7% CO2 humidified atmosphere using MCDB-131 medium with phenol red and supplemented with 15% fetal calf serum, penicillin, streptomycin, L-glutamine 10 mM, hydrocortisone 1 μg/mL, endothelial cell growth supplement (20 μg/mL).

Publication protocol

HUVECs were isolated by treatment with 0.2% type II collagenase and cultured using M-199 medium with phenol red supplemented as previously described [12]. Briefly, HUVECs were cultured at 37 °C in a 7% CO2 humidified atmosphere, in medium M-199 with phenol red and 20% fetal calf serum, penicillin, streptomycin, L-glutamine 10 mM, hydrocortisone 1 μg/mL, endothelial cell growth supplement (40 μg/mL) and heparin. The experiments were performed using pools composed of three different umbilical cords from healthy donators without personal and familiar history of cardiovascular diseases.

HMEC-1 (ATCC CRL-3243) were cultured at 37 °C in a 7% CO2 humidified atmosphere using MCDB-131 medium with phenol red and supplemented with 15% fetal calf serum, penicillin, streptomycin, L-glutamine 10 mM, hydrocortisone 1 μg/mL, endothelial cell growth supplement (20 μg/mL).

Morphology and granularity were assessed using cells without markers or stimuli using flow cytometry in a BD FACS Calibur equipment (Singapore).

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