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利用胶原酶对恒河猴血管内皮细胞进行消化,以9.92±3.34×10~3个细胞/cm~2的接种率接种于35mm培养皿中原代培养,体外培养7.7±1.82天,细胞增长了7.39±5.04倍,以1:6及1:2比例分别传第一代、第二代共历时13.89±1.36天细胞增长了147.93±88.68倍。对原代及传代细胞进行染色体及第Ⅷ因子相关抗原免疫荧光检查,证实所培养的细胞为内皮细胞。通过检测培养上清中vWF和PGF1α的含量,原代、第一代与第二代之间均无明显差异(P>0.05)。
Rhesus macaque vascular endothelial cells were digested with collagenase and inoculated into 35mm petri dishes at an inoculation rate of 9.92 ± 3.34 × 10 ~ 3 cells / cm ~ 2. The cells were cultured for 7.7 ± 1.82 days in vitro, and the cells grew by 7.39 ± 5.04-fold, respectively. The first generation was passaged at a ratio of 1: 6 and 1: 2, respectively, and the second generation showed a total increase of 147.93 ± 88.68 times at 13.89 ± 1.36 days. The primary and passaged cells were subjected to immunofluorescence staining of chromosomes and factor Ⅷ related antigens to confirm that the cultured cells were endothelial cells. There was no significant difference (P> 0.05) between the first generation and the second generation by detecting the content of vWF and PGF1α in the culture supernatant.