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Isolation and characterization of TECs and NECs.

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posted on 2014-12-01, 03:24 authored by Hitomi Ohmura-Kakutani, Kosuke Akiyama, Nako Maishi, Noritaka Ohga, Yasuhiro Hida, Taisuke Kawamoto, Junichiro Iida, Masanobu Shindoh, Kunihiko Tsuchiya, Nobuo Shinohara, Kyoko Hida

a. Cultured TECs and NECs were positive for CD31, CD105, CD144, VEGFR1, and VEGFR2 as determined by RT-PCR. TECs and NECs were negative for the monocyte marker CD11b and hematopoietic marker CD45. Human HB-EGF expression was detected in human tumor cells but not in TECs or NECs. b. Cell proliferation was measured every day for 3 days by MTS assays (*p<0.05). c. Cell migration towards VEGF was analyzed using a Boyden chamber. Cells on the filter were fixed and stained with hematoxylin after 4 h of incubation. Cells that migrated to the underside of the filter were counted (*p<0.05). d. Relative expression of VEFG-A and VEGFR2 to GAPDH in TECs and NECs was measured using real-time RT-PCR (*p<0.01).