Retinal Pigment Epithelial Cell Line with Fast Differentiation and Improved Barrier Properties (Retracted article. See vol. 14, 2022)
Retinal Pigment Epithelial Cell Line with Fast Differentiation and Improved Barrier Properties (Retracted article. See vol. 14, 2022)
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DOI：
10.3390/pharmaceutics11080412
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发表时间：
2019-08-01
期刊：
影响因子：
5.4
通讯作者：
Reinisalo, Mika
中科院分区：
文献类型：
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作者：
Hellinen, Laura;Pirskanen, Lea;Reinisalo, Mika
视网膜色素上皮（RPE）作为血液-视网膜外屏障，限制了循环中的异种抗生素进入眼睛。此外，RPE限制了玻璃体内注射药物进入循环的后路消除。因此，RPE的渗透对眼药代动力学有显著影响。RPE在老年性黄斑变性中也是一个潜在的重要药物靶点。因此，RPE细胞模型是眼科药物开发的重要工具，但缺乏可用性和可重复性问题限制了RPE细胞培养的使用。而目前应用最广泛的最佳人类细胞系ARPE19需要特殊的培养条件和较长的细胞分化时间。在本文中，我们描述了由ARPE19培养产生的细胞群，具有快速分化和改进的屏障特性。该细胞系LEPI在简单培养条件下继代培养后形成清晰的微绒毛，并迅速呈现类似rpe的鹅卵石形态。LEPI细胞具有rpe特异性功能，表达RPE65、ezrin和BEST1蛋白。在过滤器上，LEPI细胞比离体牛rpe -脉络膜形成更紧密的屏障：β受体阻滞剂（阿替洛尔、纳多洛尔、替马洛尔、品多洛尔、美托洛尔、倍他洛尔）的渗透性系数根据药物的亲脂性从0.4 × 10(-6) cm/秒到2.3 × 10(-6) cm/秒不等。这种快速分化的细胞系将是眼部研究的一项资产，因为它易于维护，生长和分化迅速，不需要专门的培养条件进行分化。因此，该细胞系适用于药物发现和开发中的小规模试验和高通量筛选。
Retinal pigment epithelium (RPE) acts as an outer blood-retinal barrier that limits the access of circulating xenobiotics to the eye. In addition, the RPE limits posterior elimination of intravitreally injected drugs to circulation. Thus, permeation in the RPE has a significant effect on ocular pharmacokinetics. The RPE is also a potentially important drug target in age-related macular degeneration. Therefore, the cell models of the RPE are important tools in ocular drug development, but poor availability and problems in reproducibility limit the use of primary RPE cell cultures. Furthermore, the best and widely used human cell line ARPE19 requires specialized culture conditions and a long time for cellular differentiation. In this paper, we describe a cell population arisen from the ARPE19 culture, with fast differentiation and improved barrier properties. This cell line, LEPI, forms clear microvilli and rapidly displays RPE-like cobblestone morphology after subculture in simple culture conditions. The LEPI cells show RPE-specific functions and expression of RPE65, ezrin, and BEST1 proteins. On filter, the LEPI cells develop tighter barrier than the ex vivo bovine RPE-choroid: permeability coefficients of beta-blockers (atenolol, nadolol, timolol, pindolol, metoprolol, betaxolol) ranged from 0.4 x 10(-6 )cm/sec to 2.3 x 10(-6) cm/sec depending on the drug lipophilicity. This rapidly differentiating cell line will be an asset in ocular studies since it is easily maintained, it grows and differentiates quickly and does not require specialized culture conditions for differentiation. Thus, this cell line is suitable for both small scale assays and high throughput screening in drug discovery and development.