原代培养人脐动脉内皮细胞的生长与增殖

THE GROWTH AND PROLIFERATION OF HUMAN UMBILICAL ARTERY ENDOTHELIAL CELLS IN PRIMARY CULTURE

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。

The human umbilical artery endothelial cells (HUAECs) were successfully cultured without using attachment agents or growth factors. HUAECs were seeded at a density of 1 × 105/cm2. The morphology,growth and proliferation of HUAECs were studied with HE staining,and their ultra-structure by transmission electron microscopy (TEM). (1) By 24h after seeding ,the HUAECs grow and form many islet-like cell clusters. By 48h,the cell clusters increase in size and can be divided into a central region with crowded endothelial cells and a peripheral region with fewer cells. By 72h,cell clusters coalesce to form incomplete monolayer. By 216h,the senescent cells shed. (2) HUAECs are relatively elongated polygonal in shape and connect with neighboring cells by short processes. The endo-plasm of HUAEC contains weak basophilic granules,while the ectoplasm is flat and thin. (3) The average cell number in a cell cluster is 17. 39±3. 79 by 24h,131. 14±22. 83 by 48h,454. 16±111. 39 by 72h respectively. These results indicate that the cell population doubled 9 times within 72h. (4) In exponential proliferating 24-72h,many amitotic figures appear but the mitotic figure is not observed. By 120h and 216h when cell growth slows down the mitotic figures appear. (5) The Weibel-Palade bodies in HUAEC are observed under TEM.