Hdmec

HUVEC (Takara Bio, Kusatsu, Japan; C-12203), HDMEC (Takara Bio; C-12212), HDBEC (Takara Bio; C-12211), and HAMON human angiosarcoma cells^{44 (link),45 (link)} (kindly provided by Riichiro Abe, Division of Dermatology, Niigata University) were cultured in Endothelial Cell Growth Medium 2 (Takara Bio; C-22111), containing 2% fetal calf serum, 5 ng/mL human epidermal growth factor, 0.2 μg/mL hydrocortisone, 22.5 μg/mL heparin, 20 ng/mL R3 insulin-like growth factor-1, 1 μg/mL ascorbic acid, 10 ng/mL human basic fibroblast growth factor, and 0.5 ng/mL VEGF. ISO-HAS-B human angiosarcoma cells (Resource Center for Biomedical Research, Institute of Development, Aging, and Cancer, Tohoku University) were cultured in DMEM (Sigma Aldrich; D6429) supplemented with 10% FBS. Cells were passaged at confluence using a Detach Kit (Takara Bio; C-41210) according to the manufacturer’s instructions, and the medium was changed every 2–3 days. The CycleavePCR Mycoplasma Detection Kit (Takara Bio; CY232) was used to test for mycoplasma contamination, and cells were confirmed to be mycoplasma free. Cell morphology was observed under a microscope and pictures were captured using a microscope camera system (Nikon).