They were washed twice with 10 ml PBS and seeded in 10 % human AB sera and antibiotics (penicillin 5 unit / ml and streptomycin 5 µg / ml and 6 µg / ml Ciprofloxacin; GIBCO - BRL, Burlington, ON) containing EGMM (
Endothelial Growth Medium Mix) made by mixing 50 % M199 (GIBCO - BRL, Burlington, ON) media in EBM - 2 media (Lonza, Walkersville, Md).