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在未用贴附基质和生长因子情况下,成功地培养了人脐动脉内皮细胞(human umbilicalartery endothelial cell,HUAEC)。接种密度为1×10~5/cm~2。采用HE染色、显微计量法及透射电镜术研究了内皮细胞的形态、生长行为与增殖。(1)接种后24h已出现岛状细胞团;48h细胞团增大,可区分出细胞密集的中央区及细胞密度较小的周边区;72h大部分细胞团已汇合;216h内皮细胞衰退、脱落。(2)HUAEC呈较长的多边形,相邻细胞间以短突相连,内胞质呈颗粒状弱嗜碱性,外胞质扁薄。(3)一个细胞团所含平均细胞数,24h为17.39±3.79个;48h为131.14±22.83个,72h为454.16±111.39个,表明在接种后72h内细胞倍增9次。(4)在细胞呈指数生长的24～72h时出现许多无丝分裂像,但未见有丝分裂像;至120h细胞生长缓慢时才出现有丝分裂像。(5)电镜下可见内皮细胞含Weibel-Palade小体。
Human umbilicalartery endothelial cells (HUAEC) were successfully cultured without attaching substrates and growth factors. Inoculation density of 1 × 10 ~ 5 / cm ~ 2. The morphology, growth behavior and proliferation of endothelial cells were studied by HE staining, microscopic measurement and transmission electron microscopy. (1) The island-shaped cell mass appeared at 24h after inoculation; the cell mass increased at 48h, the cell-dense central area and the peripheral cell with less cell density were distinguished; the majority of cell mass had confluenced at 72h; the endothelial cell receded at 216h . (2) HUAEC is a long polygon, adjacent to the short bursa connected with the cytoplasm was weakly basophilic granules, the outer cytoplasm was thin. (3) The average number of cells contained in one cell mass was 17.39 ± 3.79 at 24h, 131.14 ± 22.83 at 48h and 454.16 ± 111.39 at 72h, indicating that the cell doubling was 9 times within 72 hours after inoculation. (4) In the exponential growth of cells 24 ~ 72h appear a lot of amitosis, but no mitosis; to 120h cells only when the slow growth of mitosis. (5) Electron microscopy showed endothelial cells containing Weibel-Palade bodies.