RF/6A Chorioretinal Cells Do Not Display Key Endothelial Phenotypes.
RF/6A Chorioretinal Cells Do Not Display Key Endothelial Phenotypes.
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DOI：
10.1167/iovs.18-25215
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发表时间：
2018-12-03
影响因子：
4.4
通讯作者：
Gelfand BD
中科院分区：
文献类型：
--
作者：
Makin RD;Apicella I;Nagasaka Y;Kaneko H;Turner SD;Kerur N;Ambati J;Gelfand BD
误用不真实的细胞系被广泛认为是对生物医学科学完整性的主要威胁。鉴于大多数解决这一问题的努力都集中在DNA分析上，我们试图从解剖学、转录和功能上验证RF/6A绒毛膜细胞系，该细胞系被广泛用作内皮细胞系来模拟视网膜和脉络膜血管生成。研究人员从不同的商业经销商处获得多瓶RF/6A细胞，以验证其基因、转录组学、解剖学和功能对真正内皮细胞的保真度。无论是从头开始还是从公共数据库中获得的RF/6A细胞的转录组学特征都不符合内皮基因表达特征。与原代人内皮细胞相比，RF/6A中已建立的内皮标志物的表达非常低或无法检测到。重要的是，RF/6A细胞也没有表现出内皮细胞的功能特征，如乙酰化LDL的摄取，对TNF-α暴露的e -选择素的表达，剪切应力方向的排列，以及VEGFA刺激后AKT和ERK1/2的磷酸化。多个独立来源的RF/6A不表现出关键的内皮细胞表型。因此，这些细胞似乎不适合作为脉络膜或视网膜内皮细胞的替代品。此外，细胞系鉴定方法应扩展到基因组分析之外，包括解剖、转录和功能评估。
The misuse of inauthentic cell lines is widely recognized as a major threat to the integrity of biomedical science. Whereas the majority of efforts to address this have focused on DNA profiling, we sought to anatomically, transcriptionally, and functionally authenticate the RF/6A chorioretinal cell line, which is widely used as an endothelial cell line to model retinal and choroidal angiogenesis. Multiple vials of RF/6A cells obtained from different commercial distributors were studied to validate their genetic, transcriptomic, anatomic, and functional fidelity to bona fide endothelial cells. Transcriptomic profiles of RF/6A cells obtained either de novo or from a public data repository did not correspond to endothelial gene expression signatures. Expression of established endothelial markers were very low or undetectable in RF/6A compared to primary human endothelial cells. Importantly, RF/6A cells also did not display functional characteristics of endothelial cells such as uptake of acetylated LDL, expression of E-selectin in response to TNF-α exposure, alignment in the direction of shear stress, and AKT and ERK1/2 phosphorylation following VEGFA stimulation. Multiple independent sources of RF/6A do not exhibit key endothelial cell phenotypes. Therefore, these cells appear unsuitable as surrogates for choroidal or retinal endothelial cells. Further, cell line authentication methods should extend beyond genomic profiling to include anatomic, transcriptional, and functional assessments.
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3.1
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