Cell lines with extended in vitro growth potential from human renal proximal tubule: Characterization, response to inducers, and comparison with established cell lines
Cell lines with extended in vitro growth potential from human renal proximal tubule: Characterization, response to inducers, and comparison with established cell lines
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DOI：
10.1016/s0022-2143(97)90180-3
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发表时间：
1997-03-01
期刊：
影响因子：
--
通讯作者：
Morin, JP
中科院分区：
文献类型：
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作者：
Racusen, LC;Monteil, C;Morin, JP
用于研究人肾近曲小管上皮损伤的模型系统很少。具有延长的体外生长潜力的优化的分化人肾上皮细胞系将为原代培养或其他可用的非人哺乳动物肾细胞系提供替代模型系统。为此，从正常肾皮质中分离出人肾小管上皮细胞，并在培养物中暴露于杂交永生病毒——腺病毒12-SV40。通过有限稀释开发细胞系，并筛选三个选定的细胞系的生长模式、永生化病毒的产生、致瘤性和倍性。还监测细胞系对诱导剂和基质因子的反应，并筛选肾上皮生化特性和分化标志物的表达。这三种病毒都不是永生病毒的产生者，并且不具有致瘤性。它们以单层生长，具有中等生长动力学，并通过免疫组织化学表达肾近端肾小管上皮的标记物。它们还表现出与其他广泛使用的近端肾小管细胞系（包括 LLC-RK1、OK 和 HK-2）相当的生化特性，并且与稳定培养中的人肾小管细胞相当。含有低水平胎牛血清或表皮生长因子与甲状旁腺激素组合的生长培养基在选定的细胞系中产生最佳的生长特性、刷状缘酶表达、生化特性和葡萄糖转运。添加二甲基亚砜可以长时间维持单层形态完整。这些细胞系应该是研究人类肾近端肾小管损伤或疾病的有用模型系统。
Few model systems exist for the study of injury to human renal proximal tubule epithelium. Optimized differentiated human renal epithelial cell lines with extended in vitro growth potential would provide an alternative model system to primary culture or other available non-human mammalian kidney cell lines. For this purpose, human renal tubule epithelial cells were isolated from normal kidney cortex and exposed in culture to a hybrid immortalizing virus, adenovirus 12-SV40. Cell lines were developed by limiting dilution, and three selected cell lines were screened for growth pattern, production of immortalizing virus, tumorigenicity, and ploidy. Cell lines were also monitored for response to inducer agents and matrix factors and were screened for expression of biochemical properties and differentiation markers of renal epithelium. All three are nonproducers of the immortalizing virus and are nontumorigenic. They grow in monolayer, have intermediate growth kinetics, and express markers of renal proximal tubular epithelium by immunohistochemistry. They also express biochemical properties comparable to other widely used proximal tubular cell lines including LLC-RK1, OK, and HK-2 and comparable to human tubular cells in stable culture. Growth medium containing low levels of fetal calf serum, or epidermal growth factor combined with parathyroid hormone, produced optimal growth characteristics, brush border enzyme expression, biochemical properties, and glucose transport in a selected cell line. The addition of dimethyl sulfoxide allows maintenance in morphologically intact monolayers for prolonged periods. These cell lines should be useful model systems for the study of human renal proximal tubular injury or disease.