BRAIN CELL-DERIVED AND LIVER CELL-DERIVED FACTORS ARE REQUIRED FOR GROWTH OF HUMAN-ENDOTHELIAL CELLS IN SERUM-FREE CULTURE
BRAIN CELL-DERIVED AND LIVER CELL-DERIVED FACTORS ARE REQUIRED FOR GROWTH OF HUMAN-ENDOTHELIAL CELLS IN SERUM-FREE CULTURE
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DOI：
10.1073/pnas.81.20.6413
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发表时间：
1984-01-01
期刊：
影响因子：
--
通讯作者：
MCKEEHAN, WL
中科院分区：
文献类型：
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作者：
HOSHI, H;MCKEEHAN, WL
对超过 40 种纯化和粗制的激素和生长因子来源的促有丝分裂活性测试表明，表皮生长因子、高密度脂蛋白（牛垂体、下丘脑或全脑的提取物）以及分化人肝癌细胞条件化的培养基对培养的源自人脐静脉的内皮细胞具有促有丝分裂作用。 4 种活性剂与改进的营养培养基和胶原蛋白或纤连蛋白包被的培养表面相结合，支持内皮细胞群在无血清培养基中以 0.70-0.80 代/天的速率在低和高细胞密度下生长。脑源性活动表现出 Maciag 等人报告的特性。肝细胞衍生的活性是分化的肝癌细胞的特定产物。来自 HeLa 细胞、相对未分化的大鼠肝细胞系和人成纤维细胞的培养基没有活性。肝脏来源的纯化血浆蛋白不能替代肝癌细胞条件培养基。肝癌细胞衍生的活性是不可透析的、热不稳定的、在pH 4-11之间稳定、被胰蛋白酶和巯基乙醇处理灭活、并且在用6M尿素和苯甲基磺酰氟处理后稳定。研究结果为阐明人内皮细胞生长、功能和衰老的内分泌学提供了一个简化模型。表明神经系统和肝脏在内皮细胞再生中发挥内分泌作用。
A test of the mitogenic activity of > 40 purified and crude sources of hormones and growth factors revealed that epidermal growth factor, high density lipoprotein an extract of bovine pituitary, hypothalamus, or whole brain and the medium conditioned by differentiated human hepatoma cells were mitogenic for cultured endothelial cells derived from human umbilical vein. The 4 active agents combined with an improved nutrient medium and a collagen- or fibronectin-coated culture surface supported the growth of the endothelial cell population at a rate of 0.70-0.80 generations/day at both low and high cell densities in serum-free medium. The brain-derived activity exhibited properties reported by Maciag et al. The liver cell-derived activity was a specific product of differentiated hepatoma cells. The medium from HeLa cells, relatively undifferentiated rat liver cell lines, and human fibroblasts was inactive. Purified plasma proteins of liver origin could not substitute for the hepatoma cell-conditioned medium. The hepatoma cell-derived activity was non-dialyzable, heat-labile, stable between pH 4-11, inactivated by trypsin and mercaptoethanol treatment, and stable after treatment with 6 M urea and phenylmethylsulfonyl fluoride. The results provide a simplified model for elucidation of the endocrinology of human endothelial cell growth, function and aging. An endocrine role of both the nervous system and liver in the regeneration of endothelial cells is suggested.