論文 - 詳細
| RRC ID | 65418 |
|---|---|
| 著者 | Makin RD, Apicella I, Nagasaka Y, Kaneko H, Turner SD, Kerur N, Ambati J, Gelfand BD. |
| タイトル | RF/6A Chorioretinal Cells Do Not Display Key Endothelial Phenotypes. |
| ジャーナル | Invest Ophthalmol Vis Sci |
| Abstract |
Purpose: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. Methods: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. Results: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. Conclusions: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. |
| 巻・号 | 59(15) |
| ページ | 5795-5802 |
| 公開日 | 2018-12-3 |
| DOI | 10.1167/iovs.18-25215 |
| PII | 2717855 |
| PMID | 30508043 |
| PMC | PMC6278239 |
| MeSH | Animals Biomarkers Blotting, Western Cell Line Choroid / blood supply* E-Selectin / genetics Endothelial Cells / cytology* Endothelial Cells / metabolism Gene Expression Profiling Genetic Markers / genetics Humans Immunohistochemistry Macaca mulatta Mice Mice, Inbred C57BL Microscopy, Fluorescence Phenotype Real-Time Polymerase Chain Reaction Retinal Vessels / physiology* Transcriptome / genetics |
| IF | 3.47 |
| リソース情報 | |
| ヒト・動物細胞 | RF/6A 135(RCB1556) |