Rapid differentiation of the human RPE cell line, ARPE-19, induced by nicotinamide

Hazim, Roni A.; Volland, Stefanie; Yen, Alice; Burgess, Barry L.; Williams, David S.

doi:10.1016/j.exer.2018.10.009

Cited by 126Retinal Diseases and TreatmentsRetinal Development and DisordersConnexins and lens biology

Rapid differentiation of the human RPE cell line, ARPE-19, induced by nicotinamide

Open access

Hazim, Roni A., Volland, Stefanie, Yen, Alice, Burgess, Barry L., Williams, David S.·2019·Experimental Eye Research

DOI 10.1016/j.exer.2018.10.009PMC PMC6360117

Open this paper in CoChat — free account

Abstract

Human RPE cell lines, especially the ARPE-19 cell line, are widely-used in eye research, as well as general epithelial cell studies. In comparison with primary RPE cells, they offer relative convenience and consistency, but cultures derived from these lines are typically not well differentiated. We describe a simple, rapid method to establish cultures from ARPE-19 cells, with significantly improved epithelial cell morphology and cytoskeletal organization, and RPE-related functions. We identify the presence of nicotinamide, a member of the vitamin B family, as an essential factor in promoting the observed differentiation, indicating the importance of metabolism in RPE cell differentiation.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

Integrity signals

Registry

Crossref · 10.1016/j.exer.2018.10.009

Cites

41 of 41 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

ARPE-19 and D407 lines have genetic abnormalities, with triploidy in D407 and an abnormal karyotype in ARPE-19.
“However, these cell lines have genetic abnormalities, with triploidy in the D407 line (Davis et al., 1995), and an abnormal karyotype (most notably, a loss of one copy of chromosome 15, and an unbalanced translocation between the chromosome 15q and 19q) in the ARPE-19 line (Fasler-Kan et al., 2018; Hazim et al., 2019).”
From The cell biology of the retinal pigment epithelium (2020) · cites this paper as Hazim et al., 2019
A series of washes with trypsin selectively removes the more proliferative cells, which tend to be more loosely adhered to the substrate, leaving behind firmly attached cells that more readily form a culture with a cobblestone appearance.
“An important part of the protocol, reported by Hazim et al. for ARPE-19 cells, is a series of washes with trypsin that selectively removes the more proliferative cells, which tend to be more loosely adhered to the substrate, leaving behind firmly attached cells that more readily form a culture with a cobblestone appearance (Hazim et al., 2019).”
From The cell biology of the retinal pigment epithelium (2020) · cites this paper as Hazim et al., 2019
The pyruvate protocol for ARPE-19 differentiation required 3-4 months, whereas a more recent protocol achieved similar differentiation within 4 weeks through addition of nicotinamide instead of pyruvate.
“These protocols required 3–4 months for RPE differentiation, whereas, a more recent protocol was able to achieve a similar level of differentiation within 4 weeks through the addition of nicotinamide instead of pyruvate (Hazim et al., 2019) (Fig. 5).”
From The cell biology of the retinal pigment epithelium (2020) · cites this paper as Hazim et al., 2019

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

The Retinal Pigment Epithelium in Visual Function
Strauss, O. (2005) · Physiological Reviews · doi:10.1152/physrev.00021.2004 · Cited by 2,828 · cited 1× · see citing works
“It performs specific functions in support of the photoreceptor cells, including the reisomerization of visual chromophore, and phagocytosis of photoreceptor outer segment (POS) membrane (Strauss, 2005).”
Background
Complement factor H polymorphism and age-related macular degeneration
Edwards, A. O., Ritter, R., Abel, K. J., Manning, A., Panhuysen, C., & Farrer, L. A. (2005) · Science · doi:10.1126/science.1110189 · Cited by 2,449 · cited 5× · see citing works
“The importance of the RPE for proper vision is underscored by several retinal degenerative diseases that involve RPE pathology, including age-related macular degeneration (AMD) (Bhutto and Lutty, 2012).”
BackgroundInfluential
Complement factor H variant increases the risk of age-related macular degeneration
Haines, J. L., Hauser, M. A., Schmidt, S., Scott, W. K., Olson, L. M., Gallins, P., Spencer, K. L., Kwan, S. Y., Noureddine, M., Gilbert, J. R., et al. (2005) · Science · doi:10.1126/science.1110359 · Cited by 2,424 · cited 5× · see citing works
“The importance of the RPE for proper vision is underscored by several retinal degenerative diseases that involve RPE pathology, including age-related macular degeneration (AMD) (Bhutto and Lutty, 2012).”
BackgroundInfluential
Declining NAD+ Induces a Pseudohypoxic State Disrupting Nuclear-Mitochondrial Communication during Aging
Gomes, A. P., Price, N. L., Ling, A. J. Y., Moslehi, J. J., Montgomery, M. K., Rajman, L., White, J. P., Teodoro, J. S., Wrann, C. D., Hubbard, B. P., et al. (2013) · Cell · doi:10.1016/j.cell.2013.11.037 · Cited by 1,482 · see citing works
ARPE-19, A Human Retinal Pigment Epithelial Cell Line with Differentiated Properties
Dunn, K. C., Aotaki-Keen, A. E., Putkey, F. R., & Hjelmeland, L. M. (1996) · Experimental Eye Research · doi:10.1006/exer.1996.0020 · Cited by 1,334 · cited 1× · see citing works
“When first reported, standard cultures of the ARPE-19 cell line readily exhibited many of the characteristics of well-differentiated RPE cells, including cobblestone morphology, apical-basal polarity, and expression of RPE-specific proteins (Dunn et al., 1996).”
Background
The NAD+ Precursor Nicotinamide Riboside Enhances Oxidative Metabolism and Protects against High-Fat Diet-Induced Obesity
Cantó, C., Houtkooper, R. H., Pirinen, E., Youn, D. Y., Oosterveer, M. H., Cen, Y., Fernandez-Marcos, P. J., Yamamoto, H., Andreux, P. A., Cettour-Rose, P., et al. (2012) · Cell Metabolism · doi:10.1016/j.cmet.2012.04.022 · Cited by 1,238 · cited 1× · see citing works
“Nicotinamide has been shown to protect against oxidative stress and maintain mitochondrial health (Gomes et al., 2013; Mitchell et al., 2018), while its related compound, NR, was demonstrated to promote oxidative metabolism by means of increasing the NAD+/ NADH ratio (Canto et al., 2012).”
Background
Understanding age-related macular degeneration (AMD): Relationships between the photoreceptor/retinal pigment epithelium/Bruch’s membrane/choriocapillaris complex
Bhutto, I. A., & Lutty, G. A. (2012) · Molecular Aspects of Medicine · doi:10.1016/j.mam.2012.04.005 · Cited by 1,005 · cited 5× · see citing works
“The importance of the RPE for proper vision is underscored by several retinal degenerative diseases that involve RPE pathology, including age-related macular degeneration (AMD) (Bhutto and Lutty, 2012).”
BackgroundInfluential
Organization and execution of the epithelial polarity programme
Rodriguez-Boulan, E. J., & Macara, I. G. (2014) · Nature Reviews Molecular Cell Biology · doi:10.1038/nrm3775 · Cited by 679 · cited 1× · see citing works
“The cells typically exhibit incomplete execution of the epithelial polarity program (Rodriguez-Boulan and Macara, 2014), thus compromising their ability to represent in vivo biology.”
Background
Stem cell metabolism in tissue development and aging
Shyh‐Chang, N., Daley, G. Q., & Cantley, L. C. (2013) · Development · doi:10.1242/dev.091777 · Cited by 573 · cited 1× · see citing works
“Interestingly, many of the key enzymes and coenzymes in these metabolic pathways have been shown to become upregulated as cells differentiate into their terminal fate (Shyh-Chang et al., 2013; Hu et al., 2016).”
Background
Role of Reactive Oxygen Species in TGF-β1-Induced Mitogen-Activated Protein Kinase Activation and Epithelial-Mesenchymal Transition in Renal Tubular Epithelial Cells
Rhyu, D. Y., Yang, Y., Ha, H., Lee, G. T., Song, J. S., Uh, S., & Lee, H. B. (2005) · Journal of the American Society of Nephrology · doi:10.1681/asn.2004050425 · Cited by 523 · cited 1× · see citing works
“Interestingly, there is evidence to suggest that oxidative stress, and in particular reactive oxygen species, can promote epithelial-to-mesenchymal transition (Rhyu et al., 2005; Wang et al., 2010).”
Background
Directed differentiation of human embryonic stem cells into functional retinal pigment epithelium cells
Idelson, M., Alper, R., Obolensky, A., Ben-Shushan, E., Hemo, I., Yachimovich-Cohen, N., Khaner, H., Smith, Y., Wiser, O., Gropp, M., et al. (2009) · Cell stem cell · doi:10.1016/j.stem.2009.07.002 · Cited by 486 · cited 2× · see citing works
“Given the role that nicotinamide has been shown to play in the differentiation of RPE from pluripotent stem cells (Idelson et al., 2009; Maruotti et al., 2015; Saini et al., 2017), we tested the necessity of nicotinamide in the MEM-Nic medium for the observed morphological phenotype.”
Background
Nicotinamide Improves Aspects of Healthspan, but Not Lifespan, in Mice
Mitchell, S. J., Bernier, M., Aon, M. A., Cortassa, S., Kim, E. Y., Fang, E. F., Palacios, H. H., Ali, A., Navas-Enamorado, I., Di Francesco, A., et al. (2018) · Cell Metabolism · doi:10.1016/j.cmet.2018.02.001 · Cited by 338 · cited 1× · see citing works
“Nicotinamide has been shown to protect against oxidative stress and maintain mitochondrial health (Gomes et al., 2013; Mitchell et al., 2018), while its related compound, NR, was demonstrated to promote oxidative metabolism by means of increasing the NAD+/ NADH ratio (Canto et al.”
Background
Microtubule Organization and Function in Epithelial Cells
Müsch, A. (2003) · Traffic · doi:10.1111/j.1600-0854.2003.00149.x · Cited by 268 · cited 1× · see citing works
“In epithelial cells, the spatial organization of microtubules, in addition to their associated motors, have been shown to be critical for the polarized targeting of secretory vesicles, as well as the machinery of the endocytic pathway (Gilbert et al., 1991; Musch, 2004).”
Background
Rapid and efficient directed differentiation of human pluripotent stem cells into retinal pigmented epithelium
Buchholz, D. E., Pennington, B. O., Croze, R. H., Hinman, C. R., Coffey, P. J., & Clegg, D. O. (2013) · Stem Cells Translational Medicine · doi:10.5966/sctm.2012-0163 · Cited by 261 · see citing works
Nicotinamide-induced Mitophagy
Jang, S., Kang, H. T., & Hwang, E. S. (2012) · Journal of Biological Chemistry · doi:10.1074/jbc.m112.363747 · Cited by 259 · cited 1× · see citing works
“In particular, it serves as a precursor for the production of NAD +, NADP+, and other substrates that participate in metabolic pathways, including the TCA cycle, and the mitochondrial electron transport chain (Jang et al., 2012).”
Background
Microtubular organization and its involvement in the biogenetic pathways of plasma membrane proteins in Caco-2 intestinal epithelial cells
Gilbert, T., Le Bivic, A., Quaroni, A., & Rodriguez-Boulan, E. (1991) · The Journal of Cell Biology · doi:10.1083/jcb.113.2.275 · Cited by 234 · cited 1× · see citing works
“In epithelial cells, the spatial organization of microtubules, in addition to their associated motors, have been shown to be critical for the polarized targeting of secretory vesicles, as well as the machinery of the endocytic pathway (Gilbert et al., 1991; Musch, 2004).”
Background
Expression, secretion, and age-related downregulation of pigment epithelium-derived factor, a serpin with neurotrophic activity
Tombran‐Tink, J., Shivaram, S., Chader, G. J., Johnson, L., & Bok, D. (1995) · Journal of Neuroscience · doi:10.1523/jneurosci.15-07-04992.1995 · Cited by 218 · cited 1× · see citing works
“2F), a protein known to be secreted from the apical surface of polarized RPE cells (Tombran-Tink et al., 1995).”
Background
Nicotinamide Ameliorates Disease Phenotypes in a Human iPSC Model of Age-Related Macular Degeneration
Saini, J. S., Corneo, B., Miller, J. D., Kiehl, T. R., Wang, Q., Boles, N. C., Blenkinsop, T. A., Stern, J. H., & Temple, S. (2017) · Cell stem cell · doi:10.1016/j.stem.2016.12.015 · Cited by 187 · cited 1× · see citing works
“Given the role that nicotinamide has been shown to play in the differentiation of RPE from pluripotent stem cells (Idelson et al., 2009; Maruotti et al., 2015; Saini et al., 2017), we tested the necessity of nicotinamide in the MEM-Nic medium for the observed morphological phenotype.”
Background
Small-molecule–directed, efficient generation of retinal pigment epithelium from human pluripotent stem cells
Maruotti, J., Sripathi, S. R., Bharti, K., Fuller, J., Wahlin, K. J., Jaskula-Ranga, V., Sluch, V. M., Berlinicke, C. A., Davis, J. A., Kim, C., et al. (2015) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1422818112 · Cited by 143 · cited 1× · see citing works
“Given the role that nicotinamide has been shown to play in the differentiation of RPE from pluripotent stem cells (Idelson et al., 2009; Maruotti et al., 2015; Saini et al., 2017), we tested the necessity of nicotinamide in the MEM-Nic medium for the observed morphological phenotype.”
Background
Energy Metabolism Plays a Critical Role in Stem Cell Maintenance and Differentiation
Hu, C., Fan, L., Cen, P., Chen, E., Jiang, Z., & Li, L. (2016) · International Journal of Molecular Sciences · doi:10.3390/ijms17020253 · Cited by 138 · cited 1× · see citing works
“Interestingly, many of the key enzymes and coenzymes in these metabolic pathways have been shown to become upregulated as cells differentiate into their terminal fate (Shyh-Chang et al., 2013; Hu et al., 2016).”
Background
Functional Analysis of Serially Expanded Human iPS Cell-Derived RPE Cultures
Singh, R., Phillips, M. J., Kuai, D., Meyer, J., Martin, J. M., Smith, M. A., Perez, E. T., Shen, W., Wallace, K. A., Capowski, E. E., et al. (2013) · Investigative Ophthalmology & Visual Science · doi:10.1167/iovs.13-11943 · Cited by 136 · cited 1× · see citing works
“Furthermore, stem cell-derived RPE cells have been found to lose their polarized organization and RPE functions after only a few passages in culture (Singh et al., 2013).”
Background
Signaling Mechanism(S) of Reactive Oxygen Species in Epithelial-Mesenchymal Transition Reminiscent of Cancer Stem Cells in Tumor Progression
Wang, Z. P., Li, Y., & Sarkar, F. H. (2010) · Current Stem Cell Research & Therapy · doi:10.2174/157488810790442813 · Cited by 121 · see citing works
Retinal pigment epithelium polarity in health and blinding diseases
Caceres, P. S., & Rodriguez-Boulan, E. J. (2019) · Current Opinion in Cell Biology · doi:10.1016/j.ceb.2019.08.001 · Cited by 101 · see citing works
Induction of differentiation by pyruvate and DMEM in the human retinal pigment epithelium cell line ARPE-19
Ahmado, A., Carr, A.-J., Vugler, A. A., Semo, M., Gias, C., Lawrence, J. M., Chen, L. L., Chen, F. K., Turowski, P., da Cruz, L., et al. (2011) · Investigative Opthalmology & Visual Science · doi:10.1167/iovs.10-6374 · Cited by 100 · cited 1× · see citing works
“Although the premelanosome marker, Pmel17, has been detected in ARPE-19 cells (Ahmado et al., 2011), images of normal RPE melanosomes have never been shown.”
Background
The Retinal G Protein-coupled Receptor (RGR) Enhances Isomerohydrolase Activity Independent of Light
Wenzel, A., Oberhauser, V., Pugh, E. N., Lamb, T. D., Grimm, C., Samardzija, M., Fahl, E., Seeliger, M. W., Remé, C. E., & von Lintig, J. (2005) · Journal of Biological Chemistry · doi:10.1074/jbc.m503603200 · Cited by 92 · cited 1× · see citing works
“For western blotting, the antibodies used included: rabbit anti-RPE65 (pin 5) (Wenzel et al., 2005) (gift from Andreas Wenzel) and goat anti-FH (Quidel A312).”
Background
Microtubule motors transport phagosomes in the RPE, and lack of KLC1 leads to AMD-like pathogenesis
Jiang, M., Esteve-Rudd, J., Lopes, V. S., Diemer, T., Lillo, C., Rump, A., & Williams, D. S. (2015) · Journal of Cell Biology · doi:10.1083/jcb.201410112 · Cited by 85 · see citing works
Phenotypic Heterogeneity of Retinal Pigment Epithelial Cells In Vitro and In Situ
Burke, J. M., Skumatz, C. M. B., Irving, P. E., & McKay, B. S. (1996) · Experimental Eye Research · doi:10.1006/exer.1996.0008 · Cited by 84 · cited 1× · see citing works
“Moreover, these primary cultures exhibit significant heterogeneity among cultures from different donors (Burke et al., 1996; Kuznetsova et al., 2014).”
Background
Epigenomic and Transcriptomic Changes During Human RPE EMT in a Stem Cell Model of Epiretinal Membrane Pathogenesis and Prevention by Nicotinamide
Boles, N. C., Fernandes, M., Swigut, T., Srinivasan, R., Schiff, L., Rada-Iglesias, Á., Wang, Q., Saini, J. S., Kiehl, T. R., Stern, J. H., et al. (2020) · Stem Cell Reports · doi:10.1016/j.stemcr.2020.03.009 · Cited by 81 · see citing works
Cell Models to Study Regulation of Cell Transformation in Pathologies of Retinal Pigment Epithelium
Kuznetsova, A. V., Kurinov, A. M., & Aleksandrova, M. A. (2014) · Journal of Ophthalmology · doi:10.1155/2014/801787 · Cited by 77 · cited 1× · see citing works
“Moreover, these primary cultures exhibit significant heterogeneity among cultures from different donors (Burke et al., 1996; Kuznetsova et al., 2014).”
Background
Regulated secretion of complement factor H by RPE and its role in RPE migration
Kim, Y. H., He, S., Kase, S., Kitamura, M., Ryan, S. J., & Hinton, D. R. (2009) · Graefe s Archive for Clinical and Experimental Ophthalmology · doi:10.1007/s00417-009-1049-y · Cited by 76 · cited 1× · see citing works
“This observation is consistent with immunofluorescence data showing Factor H localization on the apical surface of human fetal RPE cultured on Transwell inserts (Kim et al., 2009).”
Result
Differentiation of RPE cells from integration-free iPS cells and their cell biological characterization
Hazim, R. A., Karumbayaram, S., Jiang, M., Dimashkie, A., Lopes, V. S., Li, D., Burgess, B. L., Vijayaraj, P., Alva-Ornelas, J. A., Zack, J. A., et al. (2017) · Stem Cell Research & Therapy · doi:10.1186/s13287-017-0652-9 · Cited by 66 · cited 2× · see citing works
“cells can be derived from embryonic and induced pluripotent stem cells (Idelson et al., 2009; Buchholz et al., 2013; Hazim et al., 2017).”
BackgroundMethod
Phagosome maturation during endosome interaction revealed by partial rhodopsin processing in retinal pigment epithelium
Wavre-Shapton, S. T., Meschede, I. P., Seabra, M. C., & Futter, C. E. (2014) · Journal of Cell Science · doi:10.1242/jcs.154757 · Cited by 60 · cited 1× · see citing works
“The ARPE-19 cells differentiated by our method establish a polarized epithelial cell model that can be used to investigate general epithelial processes, including secretory and endocytic vesicular transport, as well as RPE-specific processes such as the trafficking of phagosomes and the degradative organelles that are…”
Background
Vitronectin is responsible for serum-stimulated uptake of rod outer segments by cultured retinal pigment epithelial cells.
Miceli, M. V., Newsome, D. A., & Tate, D. J. (1997) · PubMed · Cited by 52 · see citing works
Selective Proteolysis of Arrestin by Calpain
Azarian, S. M., King, A. J., Hallett, M. A., & Williams, D. S. (1995) · Journal of Biological Chemistry · doi:10.1074/jbc.270.41.24375 · Cited by 47 · cited 1× · see citing works
“POSs were isolated using a previously published protocol for isolation of bovine POSs (Azarian et al., 1995), with a few modifications.”
Method
Defective phagosome motility and degradation in cell nonautonomous RPE pathogenesis of a dominant macular degeneration
Esteve-Rudd, J., Hazim, R. A., Diemer, T., Paniagua, A. E., Volland, S., Umapathy, A., & Williams, D. S. (2018) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1709211115 · Cited by 42 · cited 1× · see citing works
“The ARPE-19 cells differentiated by our method establish a polarized epithelial cell model that can be used to investigate general epithelial processes, including secretory and endocytic vesicular transport, as well as RPE-specific processes such as the trafficking of phagosomes and the degradative organelles that are…”
Background
Kinetics of rod outer segment phagocytosis by cultured retinal pigment epithelial cells. Relationship to cell morphology.
McLaren, M. J. (1996) · PubMed · Cited by 27 · see citing works
Rev-Erbα and Photoreceptor Outer Segments modulate the Circadian Clock in Retinal Pigment Epithelial Cells
Milićević, N., Mazzaro, N., de Bruin, I., Wils, E., ten Brink, J. B., ten Asbroek, A. L. M. A., Mendoza, J., Bergen, A. A. B., & Felder‐Schmittbuhl, M. (2019) · Scientific Reports · doi:10.1038/s41598-019-48203-3 · Cited by 26 · see citing works
The retinal pigment epithelial cell line (ARPE-19) displays mosaic structural chromosomal aberrations
Fasler-Kan, E., Aliu, N., Wunderlich, K., Ketterer, S., Ruggiero, S., Berger, S., & Meyer, P. (2018) · Methods in molecular biology · doi:10.1007/978-1-4939-7680-5_17 · Cited by 20 · cited 1× · see citing works
“The immortalization of the ARPE-19 cell line occurred spontaneously, and in a recent study (Fasler-Kan et al., 2018), it was shown that different sources of the cell line exhibited abnormal karyotypes.”
Background
Cell Culture Analysis of the Phagocytosis of Photoreceptor Outer Segments by Primary Mouse RPE Cells
Hazim, R. A., & Williams, D. S. (2018) · Methods in molecular biology · doi:10.1007/978-1-4939-7720-8_4 · Cited by 11 · cited 1× · see citing works
“A previously described double immunofluorescence labeling strategy (Hazim and Williams, 2018) was used to distinguish between POSs bound to the surface of the ARPE-19 cells and POSs that have been internalized (Fig.”
Method
Role of FGF and Hyaluronan in Choroidal Neovascularization in Sorsby Fundus Dystrophy
Wolk, A., Hatipoğlu, D., Cutler, A., Ali, M., Bell, L., Qi, J. H., Singh, R., Batoki, J. C., Karle, L., Bonilha, V. L., et al. (2020) · Cells · doi:10.3390/cells9030608 · Cited by 9 · see citing works
Effects of FTMT Expression by Retinal Pigment Epithelial Cells on Features of Angiogenesis
Buyandelger, U., Walker, D. G., Yanagisawa, D., Morimura, T., & Tooyama, I. (2020) · International Journal of Molecular Sciences · doi:10.3390/ijms21103635 · Cited by 8 · see citing works

Flagged first, then by citations

Sorted by citations

The cell biology of the retinal pigment epithelium
Lakkaraju, A., Umapathy, A., Tan, L. X., Daniele, L., Philp, N. J., Boesze-Battaglia, K., & Williams, D. S. (2020) · Progress in Retinal and Eye Research · doi:10.1016/j.preteyeres.2020.100846 · Cited by 422 · see citing works
“However, the morphological features of these cells do not appear as RPE-like as those reported by others (Ahmado et al., 2011; Hazim et al., 2019).”
BackgroundInfluential
Role of Epithelial-Mesenchymal Transition in Retinal Pigment Epithelium Dysfunction
Zhou, M., Geathers, J. S., Grillo, S. L., Weber, S. R., Wang, W., Zhao, Y., & Sundstrom, J. M. (2020) · Frontiers in Cell and Developmental Biology · doi:10.3389/fcell.2020.00501 · Cited by 199 · see citing works
Retinal pigment epithelium polarity in health and blinding diseases
Caceres, P. S., & Rodriguez-Boulan, E. J. (2019) · Current Opinion in Cell Biology · doi:10.1016/j.ceb.2019.08.001 · Cited by 101 · see citing works
Cell culture models to study retinal pigment epithelium-related pathogenesis in age-related macular degeneration
Bharti, K., den Hollander, A. I., Lakkaraju, A., Sinha, D., Williams, D. S., Finnemann, S. C., Rickman, C. B., Malek, G., & D'Amore, P. A. (2022) · Experimental Eye Research · doi:10.1016/j.exer.2022.109170 · Cited by 92 · see citing works
Epigenomic and Transcriptomic Changes During Human RPE EMT in a Stem Cell Model of Epiretinal Membrane Pathogenesis and Prevention by Nicotinamide
Boles, N. C., Fernandes, M., Swigut, T., Srinivasan, R., Schiff, L., Rada-Iglesias, Á., Wang, Q., Saini, J. S., Kiehl, T. R., Stern, J. H., et al. (2020) · Stem Cell Reports · doi:10.1016/j.stemcr.2020.03.009 · Cited by 81 · see citing works
“In addition, NAM improves cobblestone phenotype in immortalized ARPE19 cells (Hazim et al., 2019).”
Background
Polarity and epithelial-mesenchymal transition of retinal pigment epithelial cells in proliferative vitreoretinopathy
Zou, H., Shan, C., Ma, L., Liu, J., Yang, N., & Zhao, J. (2020) · PeerJ · doi:10.7717/peerj.10136 · Cited by 59 · see citing works
“Due to the importance of TJs in the maintenance of integrity and functionality of epithelial cells, several researchers have focused on novel factors that stimulate the formation of TJs, such as nicotinamide (Hazim et al., 2019) and lysophosphatidic acid (Lidgerwood et al.”
Background
Vitamin B3, nicotinamide, enhances mitochondrial metabolism to promote differentiation of the retinal pigment epithelium
Hazim, R. A., Paniagua, A. E., Tang, L., Yang, K., Kim, K. K. O., Stiles, L., Divakaruni, A. S., & Williams, D. S. (2022) · Journal of Biological Chemistry · doi:10.1016/j.jbc.2022.102286 · Cited by 40 · see citing works
“ARPE-19 cells, obtained from American Type Culture Collection (passage 19), and hfRPE from three different donors were maintained as described previously (22).”
BackgroundMethodInfluential
Reassessing the suitability of ARPE-19 cells as a valid model of native RPE biology
Pfeffer, B. A., & Fliesler, S. J. (2022) · Experimental Eye Research · doi:10.1016/j.exer.2022.109046 · Cited by 39 · see citing works
Inflammatory and cell death mechanisms induced by 7-ketocholesterol in the retina. Implications for age-related macular degeneration
Pariente, A., Peláez, R., Pérez-Sala, Á., & Larráyoz, I. M. (2019) · Experimental Eye Research · doi:10.1016/j.exer.2019.107746 · Cited by 32 · see citing works
Experimental Models to Study Epithelial-Mesenchymal Transition in Proliferative Vitreoretinopathy
Datlibagi, A., Zein-El-Din, A., Frohly, M., Willermain, F., Delporte, C., & Motulsky, E. H. (2023) · International Journal of Molecular Sciences · doi:10.3390/ijms24054509 · Cited by 31 · see citing works
“Recently, a rapid differentiation protocol using culture medium supplemented with nicotinamide has been reported to allow the cells to form a polarized epithelium with cobblestone appearance but lacking pigmentation within two to four weeks and to regain RPE functions [51].”
Background
Retinal Pigment Epithelium Under Oxidative Stress: Chaperoning Autophagy and Beyond
Markitantova, Y. V., & Simirskii, V. N. (2025) · International Journal of Molecular Sciences · doi:10.3390/ijms26031193 · Cited by 32 · see citing works
Retinal Toxicity Induced by Chemical Agents
de Araújo, D. S. M., Brito, R., Pereira‐Figueiredo, D., dos Santos‐Rodrigues, A., Logu, F. D., Nassini, R., Zin, A., & Calaza, K. D. C. (2022) · International Journal of Molecular Sciences · doi:10.3390/ijms23158182 · Cited by 30 · see citing works
“For instance, there is an established method for culturing ARPE-19 cells that uses nicotinamide to stimulate cell growth and differentiation [139].”
Method
Rethinking of TEER measurement reporting for epithelial cells grown on permeable inserts
Karakoçak, B. B., Keshavan, S., Gunasingam, G., Angeloni, S., Auderset, A., Petri‐Fink, A., & Rothen‐Rutishauser, B. (2023) · European Journal of Pharmaceutical Sciences · doi:10.1016/j.ejps.2023.106511 · Cited by 28 · see citing works
Rev-Erbα and Photoreceptor Outer Segments modulate the Circadian Clock in Retinal Pigment Epithelial Cells
Milićević, N., Mazzaro, N., de Bruin, I., Wils, E., ten Brink, J. B., ten Asbroek, A. L. M. A., Mendoza, J., Bergen, A. A. B., & Felder‐Schmittbuhl, M. (2019) · Scientific Reports · doi:10.1038/s41598-019-48203-3 · Cited by 26 · see citing works
Protection of human retinal pigment epithelial cells from oxidative damage using cysteine prodrugs
Kularatne, R. N., Bulumulla, C., Catchpole, T., Takacs, A., Christie, A. H., Stefan, M. C., & Csaky, K. G. (2020) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2020.03.024 · Cited by 26 · see citing works
Retina-on-chip: engineering functional in vitro models of the human retina using organ-on-chip technology
Gensheimer, T., Veerman, D., van Oosten, E., Segerink, L. I., Garanto, A., & van der Meer, A. D. (2025) · Lab on a Chip · doi:10.1039/d4lc00823e · Cited by 25 · see citing works
Inhibition of cellular senescence hallmarks by mitochondrial transplantation in senescence-induced ARPE-19 cells
Noh, S., Lee, S. J., Lee, T. G., Park, K., & Kim, J. H. (2022) · Neurobiology of Aging · doi:10.1016/j.neurobiolaging.2022.11.003 · Cited by 25 · see citing works
The intervention of epithelial-mesenchymal transition in homeostasis of human retinal pigment epithelial cells: a review
Gelat, B., Rathaur, P., Malaviya, P. B., Patel, B., Trivedi, K., Johar, K., & Gelat, R. (2022) · Journal of Histotechnology · doi:10.1080/01478885.2022.2137665 · Cited by 19 · see citing works
A review on eye diseases induced by blue light: pathology, model, active ingredients and mechanisms
Yuan, Y., Wu, Y., Zhao, Y., Yang, Y., An, G., Liu, Z., & Qi, D. (2025) · Frontiers in Pharmacology · doi:10.3389/fphar.2025.1513406 · Cited by 21 · see citing works
Does senescence play a role in age-related macular degeneration?
Malek, G., Campisi, J., Kitazawa, K., Webster, C. M., Lakkaraju, A., & Skowronska‐Krawczyk, D. (2022) · Experimental Eye Research · doi:10.1016/j.exer.2022.109254 · Cited by 18 · see citing works
Claudins regulate gene and protein expression of the retinal pigment epithelium independent of their association with tight junctions
Liu, F., Xu, T., Peng, S., Adelman, R. A., & Rizzolo, L. J. (2020) · Experimental Eye Research · doi:10.1016/j.exer.2020.108157 · Cited by 17 · see citing works
Metabolic reprogramming of the retinal pigment epithelium by cytokines associated with age-related macular degeneration
Hansman, D., Ma, Y., Thomas, D., Smith, J. R., Casson, R. J., & Peet, D. J. (2024) · Bioscience Reports · doi:10.1042/bsr20231904 · Cited by 18 · see citing works
“…Aldrich, St Louis, MO), 1% Glutamax (Thermo Fisher Scientific-Gibco, Grand Island, NY), 100 U/ml penicillin/streptomycin (Thermo Fisher Scientific-Gibco, Grand Island, NY) and 10 mM nicotinamide (Merck-Sigma Aldrich, St Louis, MO) to promote differentiation as previously described [85].”
Method
The CD36 Ligand‐Promoted Autophagy Protects Retinal Pigment Epithelial Cells from Oxidative Stress
Dorion, M., Mulumba, M., Kasai, S., Itoh, K., Lubell, W. D., & Ong, H. (2021) · Oxidative Medicine and Cellular Longevity · doi:10.1155/2021/6691402 · Cited by 17 · see citing works
“ARPE-19 cells (American Type Cell Collection) were cultured in complete medium and differentiated as previously described with slight modifications [29].”
Mitochondrial transplantation attenuates oligomeric amyloid-beta-induced mitochondrial dysfunction and tight junction protein destruction in retinal pigment epithelium
Noh, S., Lee, S. J., Cho, C. S., Jo, D. H., Park, K., & Kim, J. H. (2023) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2023.12.012 · Cited by 16 · see citing works
ITF2357 regulates NF‐κB signaling pathway to protect barrier integrity in retinal pigment epithelial cells
Lim, R. R., Mahaling, B., Tan, A., Mehta, M., Kaur, C., Hunziker, W., Kim, J. E., Barathi, V. A., Ghosh, A., & Chaurasia, S. S. (2024) · The FASEB Journal · doi:10.1096/fj.202301592r · Cited by 15 · see citing works
“22,23 Since inflammation is a critical feature in DME, pro-inflammatory cytokine TNF α was introduced to simulate stress in the cellular environment.”
Background
In vitro anti-inflammatory activity and molecular docking of Peperomia pellucida (L.) Kunth extract via the NF-κB and PPAR-γ signalling in human retinal pigment epithelial cells
Ho, K. L., Yong, P. H., Wang, C. W., Lim, S., Kuppusamy, U. R., Arumugam, B., Ngo, C. T., & Ng, Z. X. (2024) · Bioorganic Chemistry · doi:10.1016/j.bioorg.2024.107969 · Cited by 15 · see citing works
ARP-T1-associated Bazex–Dupré–Christol syndrome is an inherited basal cell cancer with ciliary defects characteristic of ciliopathies
Park, H., Papanastasi, E., Blanchard, G., Chiticariu, E., Bachmann, D., Plomann, M., Morice‐Picard, F., Vabres, P., Smahi, A., Huber, M., et al. (2021) · Communications Biology · doi:10.1038/s42003-021-02054-9 · Cited by 16 · see citing works
Dopamine receptor signaling regulates fibrotic activation of retinal pigmented epithelial cells
Gao, A. Y., Link, P. A., Bakri, S. J., & Haak, A. J. (2022) · American Journal of Physiology-Cell Physiology · doi:10.1152/ajpcell.00468.2021 · Cited by 13 · see citing works
Expression of two major isoforms of MYO7A in the retina: Considerations for gene therapy of Usher syndrome type 1B
Gilmore, W. B., Hultgren, N. W., Chadha, A., Barocio, S. B., Zhang, J., Kutsyr, O., Flores‐Bellver, M., Canto-Soler, M. V., & Williams, D. S. (2023) · Vision Research · doi:10.1016/j.visres.2023.108311 · Cited by 12 · see citing works
In Vitro Models of Diabetes: Focus on Diabetic Retinopathy
Galgani, G., Bray, G., Martelli, A., Calderone, V., & Citi, V. (2024) · Cells · doi:10.3390/cells13221864 · Cited by 12 · see citing works
Effects of E-Cigarette Refill Liquid Flavorings with and without Nicotine on Human Retinal Pigment Epithelial Cells: A Preliminary Study
Goenka, S., & Simon, S. R. (2021) · International Journal of Environmental Research and Public Health · doi:10.3390/ijerph182111655 · Cited by 13 · see citing works
“Although ARPE-19 can be differentiated and fully polarized with melanization after 4 months of culture [74,75] using a specialized medium with 1% serum or in a shorter span of 1 week by supplementation of exogenous chemical, nicotinamide, to induce differentiation [76], the use of undifferentiated ARPE-19 cells is a r…”
Result
Fucoxanthin Pretreatment Ameliorates Visible Light-Induced Phagocytosis Disruption of RPE Cells under a Lipid-Rich Environment via the Nrf2 Pathway
Liu, Y., Guo, Z., Wang, S., Liu, Y., & Wei, Y. (2021) · Marine Drugs · doi:10.3390/md20010015 · Cited by 12 · see citing works
“Construction of Differentiated RPE Cell Monolayer The differentiated RPE cell monolayer was cultured as described in previous studies [39,40].”
Method
Magnetically Controllable and Degradable Milliscale Swimmers as Intraocular Drug Implants
Yıldız, E., Bozuyuk, U., Yildiz, E., Wang, F., Han, M., Karacakol, A. C., Sheehan, D., Yu, Y., & Sitti, M. (2025) · Advanced Science · doi:10.1002/advs.202507569 · Cited by 12 · see citing works
The versatile roles of retinal pigment epithelium in the pathophysiology of retinitis pigmentosa
Ghanawi, H., & Koch, S. (2025) · Progress in Retinal and Eye Research · doi:10.1016/j.preteyeres.2025.101390 · Cited by 13 · see citing works
Association of Different ABO and Rh Blood Groups with the Erythroprotective Effect of Extracts from Navicula incerta and Their Anti-Inflammatory and Antiproliferative Properties
Ruíz‐Cruz, S., González-Vega, R. I., Robles‐Zepeda, R. E., Reyes‐Díaz, A., López‐Elías, J. A., Álvarez-Ainza, M. L., Cinco-Moroyoqui, F. J., Moreno-Corral, R. A., Wong‐Corral, F. J., Borboa‐Flores, J., et al. (2022) · Metabolites · doi:10.3390/metabo12121203 · Cited by 11 · see citing works
“with the typical structural morphology of healthy cells [76].”
Background
Retinal Pigment Epithelium Cell Culture
Klettner, A. (2020) · Springer eBooks · doi:10.1007/978-3-030-28384-1_17 · Cited by 11 · see citing works
Development of a CRISPRi Human Retinal Pigmented Epithelium Model for Functional Study of Age-Related Macular Degeneration Genes
Wang, J.-H., Urrutia-Cabrera, D., Lees, J. G., Mora, S. M., Nguyen, T., Hung, S. S. C., Hewitt, A. W., Lim, S. Y., Edwards, T. L., & Wong, R. C. B. (2023) · International Journal of Molecular Sciences · doi:10.3390/ijms24043417 · Cited by 10 · see citing works
“Using an established method to induce polarisation of ARPE19 with nicotinamide treatment [21], we verified that ARPE19-KRAB retained the ability to form a monolayer with organised hexagonal RPE morphology, as shown by expression of the tight junction protein ZO-1 (Figure 1C).”
BackgroundMethod
Type I and III interferons shape the retinal cytokine network and barrier function in an in vitro model of ocular toxoplasmosis
Geiller, B., Greigert, V., Hillenbrand, C. A., Gommenginger, C., Beal, L., Brunet, J., Filisetti, D., Villard, O., Denis, J., & Pfaff, A. W. (2023) · Frontiers in Immunology · doi:10.3389/fimmu.2023.1148037 · Cited by 10 · see citing works
“The use of an established differentiation protocol (20) allowed us to approach primary RPE cell characteristics, like the expression of recognized markers and the ZO-1 localization to tight junctions.”
Background
In vitro laboratory models of proliferative vitreoretinopathy
Gao, A. Y., Haak, A. J., & Bakri, S. J. (2023) · Survey of Ophthalmology · doi:10.1016/j.survophthal.2023.05.007 · Cited by 10 · see citing works
Cell sheet engineering platforms integrating antibacterial and thermo-responsive functionalities: Cu-nanoparticle-loaded P4VP brushes for retinal cell sheet harvesting
Cieślik, A., Shymborska, Y., Tymetska, S., Stetsyshyn, Y., Bernasik, A., Brzychczy‐Włoch, M., Drożdż, K., Szajna, K., Krok, F., Budkowski, A., et al. (2025) · Chemical Engineering Journal · doi:10.1016/j.cej.2025.162985 · Cited by 9 · see citing works
Maillard Reaction Crosslinked Alginate-Albumin Scaffolds for Enhanced Fenofibrate Delivery to the Retina: A Promising Strategy to Treat RPE-Related Dysfunction
Zadeh, M. A., Alany, R. G., Satarian, L., Shavandi, A., Almousa, M. A., Brocchini, S., & Khoder, M. (2023) · Pharmaceutics · doi:10.3390/pharmaceutics15051330 · Cited by 9 · see citing works
“This cell line is commonly used for eye research applications due to its convenience and consistency for cell culture studies [64].”
Background
Role of FGF and Hyaluronan in Choroidal Neovascularization in Sorsby Fundus Dystrophy
Wolk, A., Hatipoğlu, D., Cutler, A., Ali, M., Bell, L., Qi, J. H., Singh, R., Batoki, J. C., Karle, L., Bonilha, V. L., et al. (2020) · Cells · doi:10.3390/cells9030608 · Cited by 9 · see citing works
“720,000 cells and 100,000 cells were plated per well in each well of a 12-well plate or 24-well plate, respectively using a previously published protocol [36].”
Method
New Technologies to Study Functional Genomics of Age-Related Macular Degeneration
Nguyen, T., Urrutia-Cabrera, D., Liou, R. H.-C., Luu, C. D., Guymer, R. H., & Wong, R. C. B. (2021) · Frontiers in Cell and Developmental Biology · doi:10.3389/fcell.2020.604220 · Cited by 9 · see citing works
Improvement of human embryonic stem cell-derived retinal pigment epithelium cell adhesion, maturation, and function through coating with truncated recombinant human vitronectin
Zhu, X., Zhang, T., Liu, S., Bai, X., Huang, X., Jiang, M., & Sun, X. (2021) · International Journal of Ophthalmology · doi:10.18240/ijo.2021.08.04 · Cited by 9 · see citing works
NAD+ metabolism and eye diseases: current status and future directions
Liu, S., & Zhang, W. (2023) · Molecular Biology Reports · doi:10.1007/s11033-023-08692-y · Cited by 7 · see citing works
“Researchers found that NAM induced rapid differen - tiation of ARPE-19 cells, which was reversed by removing supplemented NAM [ 69].”
Background
Extracellular vesicles from retinal pigment epithelial cells expressing R345W‐Fibulin‐3 induce epithelial‐mesenchymal transition in recipient cells
Zhou, M., Zhao, Y., Weber, S. R., Gates, C., Carruthers, N. J., Chen, H., Liu, X., Wang, H., Ford, M., Swulius, M. T., et al. (2023) · Journal of Extracellular Vesicles · doi:10.1002/jev2.12373 · Cited by 8 · see citing works
“Though ARPE-19 cells were cultured to confluence so that a monolayer and tight junction had formed, the RPE characteristics of these cells could be improved if they had been left to differentiate for 2 weeks with nicotinamide (Hazim et al., 2019).”
Background
Buspirone Enhances Cell Survival and Preserves Structural Integrity during Oxidative Injury to the Retinal Pigment Epithelium
Biswal, M. R., Paulson, R. J., Vichare, R., & Lewin, A. S. (2023) · Antioxidants · doi:10.3390/antiox12122129 · Cited by 8 · see citing works
“Following di ﬀ erentiation [26], ARPE-19 cells were subjected to oxidative stress induced using paraquat (1000 µM) treatment. employed ZO-1 immunocytochemistry on cells cultured in 8-well chamber slides.”
BackgroundMethodInfluential
High‐Throughput Microfluidic 3D Outer Blood‐Retinal Barrier Model in a 96‐Well Format: Analysis of Cellular Interactions and Barrier Function in Retinal Health and Disease
Kim, J., Song, Y., Jolly, A. L., Hwang, T., Kim, S., Lee, B., Jang, J., Jo, D. H., Baek, K., Liu, T., et al. (2024) · Advanced Materials Technologies · doi:10.1002/admt.202400634 · Cited by 8 · see citing works
Mesenchymal Stem Cell Secretome Enhancement by Nicotinamide and Vasoactive Intestinal Peptide: A New Therapeutic Approach for Retinal Degenerative Diseases
Alonso-Alonso, M. L., Srivastava, G. K., Usategui‐Martín, R., García-Gutiérrez, M. T., Jimeno, J. C. P., & Fernandez‐Bueno, I. (2020) · Stem Cells International · doi:10.1155/2020/9463548 · Cited by 8 · see citing works
“Besides, NIC has been used to induce the differentiation of ARPE-19 cells [38].”
Method
A Novel Method of Mouse RPE Explant Culture and Effective Introduction of Transgenes Using Adenoviral Transduction for In Vitro Studies in AMD
Shang, P., Stepicheva, N. A., Liu, H., Chowdhury, O., Franks, J. M., Sun, M., Hose, S., Ghosh, S., Yazdankhah, M., Strizhakova, A., et al. (2021) · International Journal of Molecular Sciences · doi:10.3390/ijms222111979 · Cited by 8 · see citing works
“However, the limitations of this technique are low transepithelial resistance, lack of pigmentation, and abnormal karyotype [4].”
Background
Effects of FTMT Expression by Retinal Pigment Epithelial Cells on Features of Angiogenesis
Buyandelger, U., Walker, D. G., Yanagisawa, D., Morimura, T., & Tooyama, I. (2020) · International Journal of Molecular Sciences · doi:10.3390/ijms21103635 · Cited by 8 · see citing works
“ARPE-19 cells were differentiated according to a previously published method with modifications [37].”
BackgroundMethod
Metabolic Phenotyping of Healthy and Diseased Human RPE Cells
Rizwan, S., Toothman, B., Li, B., Engel, A. L., Lim, R. R., Niernberger, S., Lu, J., Ratliff, C., Xiang, Y., Eminhizer, M., et al. (2024) · Investigative Ophthalmology & Visual Science · doi:10.1167/iovs.65.11.5 · Cited by 7 · see citing works
Retinal Pigment Epithelium and Monocytes’ Mitochondrial Control of Ferroptosis and its Relevance to Age-Related Macular Degeneration
Singh, M. K., Singh, L., Atilano, S. R., Chwa, M., Salimiaghdam, N., & Kenney, M. C. (2025) · Molecular Neurobiology · doi:10.1007/s12035-025-04832-6 · Cited by 7 · see citing works
“We utilized the method used by Hazim et al. to efficiently differentiate ARPE-19 cells in vitro [26, 27].”
Injectable Hydrogel Guides Neurons Growth with Specific Directionality
Tseng, Y., Ma, T.-L., Tan, D.-H., Su, A.-J. A., Washington, K. M., Wang, C., Huang, Y., Wu, M., & Su, W. (2023) · International Journal of Molecular Sciences · doi:10.3390/ijms24097952 · Cited by 7 · see citing works
Exploring the therapeutic potential of MOTS-c in age-related macular degeneration: from cellular responses to patient-derived cybrids
Mohtashami, Z., Schneider, K., Azimi, R., Atilano, S. R., Chwa, M., Kenney, M. C., & Singh, M. K. (2025) · Human Cell · doi:10.1007/s13577-025-01188-w · Cited by 6 · see citing works
“MEM-Nic media contained MEM alpha with GlutaMAX, 1% FBS, 1% Penicillin/Streptomycin, 1% N1 supplement (a specific cell culture supplement to promote the differentiation of neural cells), taurine (0.25 mg/ml), hydrocortisone (20 ng/ml), triiodo-thyronine (0.013 ng/ml), and 10 mM nicotinamide [31].”
Influential
Tissue engineering of outer blood retina barrier for therapeutic development
Hampton, C., Bharti, K., & Song, M. J. (2024) · Current Opinion in Biomedical Engineering · doi:10.1016/j.cobme.2024.100538 · Cited by 6 · see citing works
Effects of galectin-3 protein on UVA-induced damage in retinal pigment epithelial cells
Andrade, F. E. C., Correia-Silva, R. D., Covre, J. L., Lice, I., Gomes, J. Á. P., & Gil, C. D. (2022) · Photochemical & Photobiological Sciences · doi:10.1007/s43630-022-00294-0 · Cited by 6 · see citing works
“Although it has been shown that ARPE-19 do express various proteins and phenotype that are relevant for the RPE phenotype [29, 30], literature search indicates that more predictive preclinical in vitro models are required when working with cell line, such as ARPE-19 [31].”
Background
The Antioxidant Tetrapeptide Epitalon Enhances Delayed Wound Healing in an in Vitro Model of Diabetic Retinopathy
Gatta, M., Dovizio, M., Milillo, C., Ruggieri, A. G., Sallese, M., Antonucci, I., Trofimov, A. V., Khavinson, V., Trofimova, S., Bruno, A., et al. (2025) · Stem Cell Reviews and Reports · doi:10.1007/s12015-025-10911-x · Cited by 5 · see citing works
“These results are consistent are a common model for studying retinal function [68, 69], their standard culture medium already contains elevated glucose levels higher than physiological norms.”
Evaluation of mesenchymal stem cells as an in vitro model for inherited retinal diseases
Dodina, M., Gurtsieva, D., Karabelsky, A. V., & Minskaia, E. (2024) · Frontiers in Cell and Developmental Biology · doi:10.3389/fcell.2024.1455140 · Cited by 5 · see citing works
KIF11 Inhibition Induces Retinopathy Progression by Affecting Photoreceptor Cell Ciliogenesis and Cell Cycle Regulation in Development
Xu, Y., Chen, J., Wang, X., Huang, M., Wei, X., Luo, X., Wei, Y., & She, Z. (2025) · Advanced Biology · doi:10.1002/adbi.202400748 · Cited by 5 · see citing works
Nicotinamide restricts neural precursor proliferation to enhance catecholaminergic neuronal subtype differentiation from mouse embryonic stem cells
Griffin, S. M., Pickard, M. R., Hawkins, C., Williams, A. C., & Fricker, R. A. (2020) · PLoS ONE · doi:10.1371/journal.pone.0233477 · Cited by 5 · see citing works
Tissue Inhibitor of Metalloproteinase 3 (TIMP3) mutations increase glycolytic activity and dysregulate glutamine metabolism in RPE cells
Grenell, A., Singh, C., Raju, M., Wolk, A., Dalvi, S., Jang, G.-F., Crabb, J. S., Hershberger, C. E., Manian, K. V., Hernandez, K. N., et al. (2024) · Molecular Metabolism · doi:10.1016/j.molmet.2024.101995 · Cited by 5 · see citing works
“ARPE-19 cells were cultured using a method previously shown to increase epithelial cell morphology, cytoskeletal organization and RPE-related functions [42].”
Method
Modeling Choroideremia Disease with Isogenic Induced Pluripotent Stem Cells
Fonseca, A. F., Coelho, R., Lopes‐da‐Silva, M., de Lemos, L., Hall, M. J., Oliveira, D., Falcão, A. S., Tenreiro, S., Seabra, M. C., & Antas, P. (2024) · Stem Cells and Development · doi:10.1089/scd.2024.0105 · Cited by 5 · see citing works
“12,13 In addition, ARPE-19 cells produce some RPE-speci ﬁ c mRNA transcripts without the corresponding protein expression, such as RPE65, BEST1, and CLDN19, leading to an incompletely differentiated phenotype.”
Epigenetic Modifications in the Retinal Pigment Epithelium of the Eye During RPE-Related Regeneration or Retinal Diseases in Vertebrates
Grigoryan, E. N., & Markitantova, Y. V. (2025) · Biomedicines · doi:10.3390/biomedicines13071552 · Cited by 4 · see citing works
Influence of Substrate Stiffness on iPSC-Derived Retinal Pigmented Epithelial Cells
Wendland, R. J., Tucker, B. A., & Worthington, K. S. (2024) · Stem Cells Translational Medicine · doi:10.1093/stcltm/szae022 · Cited by 4 · see citing works
Peperomia pellucida (L.) Kunth suppresses glycation‐induced inflammatory response in human retinal pigment epithelial cell line ARPE‐19 via JAK‐STAT3 signaling
Ho, K. L., Yong, P. H., Lim, S., & Ng, Z. X. (2024) · Archiv der Pharmazie · doi:10.1002/ardp.202400299 · Cited by 4 · see citing works
Sulindac prevents increased mitochondrial VDAC1 expression and cell surface mistargeting induced by pathological conditions in retinal cells
Mohammad, T., Sjögren, M., & Salehi, A. A. (2024) · Biochemical and Biophysical Research Communications · doi:10.1016/j.bbrc.2024.150558 · Cited by 4 · see citing works
Epithelial-Mesenchymal Transition: Molecular Mechanisms of Retinal Pigment Epithelial Cell Activation
Kuznetsova, A. (2022) · Russian Journal of Developmental Biology · doi:10.1134/s1062360422060054 · Cited by 3 · see citing works
“Recently, the role of nicotinamide in the return of RPE cells from a dedifferentiated with EMT state to a differentiated state was closely studied in vitro (Hazim et al., 2019; Boles et al., 2020; Zhou et al., 2020).”
Background
Machine learning-based 3D segmentation of mitochondria in polarized epithelial cells
Hultgren, N. W., Zhou, T., & Williams, D. S. (2024) · Mitochondrion · doi:10.1016/j.mito.2024.101882 · Cited by 4 · see citing works
Intracellular Trafficking Pathway and ROS Scavenging Activity of Glycol Chitosan‐Coated Cerium Oxide Nanoparticles in a Pathological AMD‐Like Model
Kwon, Y., & Han, Z. (2024) · Advanced Therapeutics · doi:10.1002/adtp.202400030 · Cited by 4 · see citing works
Isolation of Primary Porcine Retinal Pigment Epithelial Cells for In Vitro Modeling
Paterson, C. A., Weatherston, D., Teeples, T., & Vargis, E. (2024) · Journal of Visualized Experiments · doi:10.3791/66079 · Cited by 3 · see citing works
Interferons and Toxoplasma gondii shape PD-L1 regulation in retinal barrier cells: the critical role of proteases
Geiller, B., Cevallos, C., Tsybenko, I., Arnoux, L., Felder‐Schmittbuhl, M., & Pfaff, A. W. (2025) · Frontiers in Immunology · doi:10.3389/fimmu.2025.1607247 · Cited by 3 · see citing works
“Human RPE ARPE-19 cells were differentiated according to the protocol described by Hazim et al. (12).”
AAV-mediated multiple gene therapy combining VEGFA-targeting miR-agshRNAs and PEDF for the suppression of choroidal neovascularization
Fabian-Jessing, B. K., Askou, A. L., Jakobsen, T. S., Adsersen, R. L., Lindholm, A. B., Bjerre, A. K. K., Alsing, S., Bek, T., Aagaard, L., & Corydon, T. J. (2026) · Molecular Therapy — Nucleic Acids · doi:10.1016/j.omtn.2026.102833 · Cited by 3 · see citing works
Accelerated maturation of ARPE ‐19 cells for the translational assessment of gene therapy
Bernd, J., Plastino, F., Karayannis, J. J., Kvanta, A., Locri, F., & André, H. (2024) · The FASEB Journal · doi:10.1096/fj.202301707rr · Cited by 3 · see citing works
“12,13 In addition, several laminins occur naturally in the multilayered Bruch's membrane with laminin 521, 511, 332, and 111 as the main isoforms expressed.”
Metabolomic signature in ocular dosing: Exploring the metabolic impacts of sublethal high-dose naringenin on ARPE-19 cells
Kaplan, O., Koç, E., Türk, S., Küçükkılınç, T. T., Göktaş, Z., & Çelebi̇er, M. (2024) · European Journal of Integrative Medicine · doi:10.1016/j.eujim.2024.102414 · Cited by 3 · see citing works
Tauroursodeoxycholic Acid Confers Protection Against Oxidative Stress via Autophagy Induction in Retinal Pigment Epithelial Cells
Zubieta, D., Warden, C., Bhattacharya, S., & Brantley, M. A. (2025) · Current Issues in Molecular Biology · doi:10.3390/cimb47040224 · Cited by 3 · see citing works
“TUDCA’s ability to induce autophagy flux was observed in both ARPE-19 cells, a human RPE cell line commonly used for research [42], and iPSC-derived RPE cells, which are derived from pluripotent stem cells and more closely mimic the native RPE’s characteristics [43].”
Complement C3 Activation in the Human Retinal Pigment Epithelium
Schikora, J., Nickel, A., Bergert, J., Hähnel, R., Dort, A., King, B. C., Schayan-Araghi, S. Y., Banerjee, P., Wolf, H., May‐Simera, H., et al. (2025) · Investigative Ophthalmology & Visual Science · doi:10.1167/iovs.66.9.67 · Cited by 3 · see citing works
“55 Application of HM protocols, as described by Hazim et al., 37 partially improved epithelial traits.”
The Silk Protein Sericin Promotes Viability of ARPE-19 and Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelial Cells in vitro
Khan, A. Z., Utheim, T. P., Moe, M. C., Aass, H. C. D., Sapkota, D., Vallenari, E. M., & Eidet, J. R. (2020) · Current Eye Research · doi:10.1080/02713683.2020.1809001 · Cited by 3 · see citing works
The GPCR Antagonistic Drug CM-20 Stimulates Mitochondrial Activity in Human RPE Cells
Chang, Q., Chen, S., & Yang, T. (2022) · The Open Biochemistry Journal · doi:10.2174/1874091x-v16-e2206270 · Cited by 3 · see citing works
Spatiotemporal Coordination of RPE Cell Quality by Extracellular Vesicle miR-494-3p Via Competitive Interplays With SIRT3 or PTEN
Hamuro, J., Yamashita, T., Otsuki, Y., Hiramoto, N., Adachi, M., Miyatani, T., Tanaka, H., Ueno, M., Kinoshita, S., & Sotozono, C. (2023) · Investigative Ophthalmology & Visual Science · doi:10.1167/iovs.64.5.9 · Cited by 3 · see citing works
“Hazim et al. 27 recently reported that during NAM-induced differentiation, the mitochondria became larger with more tightly folded The PTEN versus SIRT3 balance in mitochondria may be critical as one of the causal elements in the degeneration of RPE cells.”
Influential
Clustered ARPE-19 cells distinct in mitochondrial membrane potential may play a pivotal role in cell differentiation
Miyatani, T., Tanaka, H., Numa, K., Uehara, A., Otsuki, Y., Hamuro, J., Kinoshita, S., & Sotozono, C. (2024) · Scientific Reports · doi:10.1038/s41598-024-73145-w · Cited by 3 · see citing works
“Cell morphology was monitored periodically by culturing ARPE-19 cells in standard medium (DMEM/F12) or differentiation medium (MEM-Nic) for 4 weeks as previously reported 17,21 .”
Influential
Productive infection of the retinal pigment epithelium by SARS-CoV-2: Initial effects and consideration of long-term consequences
Hultgren, N. W., Petcherski, A., Torriano, S., Komirisetty, R., Sharma, M. B., Zhou, T., Burgess, B. L., Ngo, J., Osto, C. A., Shabane, B., et al. (2024) · PNAS Nexus · doi:10.1093/pnasnexus/pgae500 · Cited by 2 · see citing works
Regulation of RPE65 expression in human retinal pigment epithelium cells
Postnikova, O. A., Samuel, W., Uppal, S., Bernstein, S. L., Poliakov, E., Rogozin, I. B., & Redmond, T. M. (2025) · Scientific Reports · doi:10.1038/s41598-025-12926-3 · Cited by 2 · see citing works
“We adopted the RPE differentiation protocol of Hazim et al. 9 .”
Safety and effectiveness of vital dyes for intraocular surgery
Romano, M. R., Ferrara, M., Szurman, P., Spadaro, A., Levis, H. J., Govetto, A., Rossi, T., Platania, C. B. M., Januschowski, K., Bucolo, C., et al. (2025) · Progress in Retinal and Eye Research · doi:10.1016/j.preteyeres.2025.101412 · Cited by 2 · see citing works
A TGFB2/TNF-induced in vitro model of proliferative vitreoretinopathy (PVR) using ARPE-19 cells confirms nicotinamide as an inhibitor of EMT and VEGFA secretion
Huang, Y., Meister, R., Lindziute, M., Binter, M., Tode, J., Framme, C., & Fuchs, H. R. (2026) · PLoS ONE · doi:10.1371/journal.pone.0340614 · Cited by 2 · see citing works
Sigma-2 receptor modulators alter low-density lipoprotein receptor-mediated lipid uptake in retinal pigment epithelial cells
Lizama, B. N., Reaver, A., Look, G. C., Caggiano, A. O., & Hamby, M. E. (2026) · Experimental Eye Research · doi:10.1016/j.exer.2026.110944 · Cited by 2 · see citing works
Retinal pigment epithelium exhibits gene expression and phagocytic activity alterations when exposed to retinoblastoma chemotherapeutics
Cerna-Chavez, R., Rozanska, A. L., Poretti, G. L., Benvenisty, N., Parulekar, M., & Lako, M. (2023) · Experimental Eye Research · doi:10.1016/j.exer.2023.109542 · Cited by 2 · see citing works
Using Small Molecules to Reprogram RPE Cells in Regenerative Medicine for Degenerative Eye Disease
Rzhanova, L. A., Алпеева, Е. В., & Aleksandrova, M. A. (2024) · Cells · doi:10.3390/cells13231931 · Cited by 2 · see citing works
Investigating the Effect of Esterification on Retinal Pigment Epithelial Uptake Using Rhodamine B Derivatives
Bulumulla, C., Kularatne, R. N., Catchpole, T., Takacs, A., Christie, A. H., Gilfoyle, A., Nguyen, T. D., Stefan, M. C., & Csaky, K. G. (2020) · Translational Vision Science & Technology · doi:10.1167/tvst.9.6.18 · Cited by 2 · see citing works
Reprogramming Factors Activate a Non-Canonical Oxidative Resilience Pathway That Can Rejuvenate RPEs and Restore VisionPreprint
Lu, Y. R., Cameron, J. C., Hu, Y., Shen, H., Shirahama, S., Tyshkovskiy, A., Chen, Z., Ai, J., Zhu, D. Y., Karg, M. M., et al. (2025) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2025.08.30.673239 · Cited by 2 · see citing works
Age-Related Macular Degeneration and its Current Treatment Strategies: An Updated Review
Gelat, B., Trivedi, K., Malaviya, P. B., Rathaur, P., Patel, B., Gelat, R., & Johar, K. (2024) · IntechOpen eBooks · doi:10.5772/intechopen.1004075 · Cited by 1 · see citing works
A novel accessible in vitro model of proliferative vitreoretinal diseases shows facilitated epithelial mesenchymal transition through aquaporin-1
Datlibagi, A., Duveau, C., Zein-El-Din, A., Bolaky, N., Lambert, V., Perret, J., Willermain, F., Delporte, C., & Motulsky, E. H. (2025) · Scientific Reports · doi:10.1038/s41598-025-28041-2 · Cited by 1 · see citing works
“…to the use of ARPE-19 cells due to their lack of typical RPE phenotype and functionality may be alleviated using a differentiation protocol recently described, involving nicotinamide supplementation in the culture medium to obtain mature RPE showing a cobblestone morphology within a few weeks 13 .”
The contribution of ferroptosis to the epithelial-mesenchymal transition phenotype in models of age-related macular degeneration
Yang, X., Xu, W., Xie, H., Guo, X., Zhu, F., Xing, Y., & Chen, C. (2026) · Frontiers in Cell and Developmental Biology · doi:10.3389/fcell.2026.1718715 · Cited by 1 · see citing works
Neuropilin-2 Deficiency Promotes Mitochondrial Dysfunction and NAD⁺-Dependent Cellular Senescence in Retinal Degeneration
Ou, L., Cui, B., Zhou, L., Liu, K., Shi, L., Zhou, T. G., Zeng, L., Zhou, H., Xu, H., Zheng, Y., et al. (2026) · Investigative Ophthalmology & Visual Science · doi:10.1167/iovs.67.3.11 · Cited by 1 · see citing works
Genome-wide CRISPR knockout screening identifies novel disease-associated genes in retinal pigment epithelium cells
Li, R., Xie, L., Hu, J., Liu, B., Zhang, H., & Qian, H. (2026) · Experimental Eye Research · doi:10.1016/j.exer.2026.111032 · Cited by 1 · see citing works
A critical role for mitochondrial dynamics in cigarette smoke condensate-induced RPE senescence
Zhou, Q., Zhang, Z., Wu, Y. J., Dong, C., Wu, C.-C., Handa, J. T., Ma, J., & Wang, S. (2026) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2026.06.060 · Cited by 1 · see citing works
Investigating the effects of 7‐ketocholesterol on retinal pigment epithelium bioenergetics
Dey, S., Catchpole, T., Takacs, A., & Csaky, K. G. (2023) · The FASEB Journal · doi:10.1096/fj.202300101r · Cited by 1 · see citing works
Potential role of CTSS in AMDImmune modulatory and anti-angiogenic effects of cathepsin S knockdown in ARPE-19 cells
Preya, U. H., Sayed, S., Nguyen, N., & Kim, J. T. (2024) · Experimental Eye Research · doi:10.1016/j.exer.2024.109981 · Cited by 1 · see citing works
MicroRNA Regulation for Inflammasomes in High Glucose‐Treated ARPE‐19 Cells
Kim, J. H., Yu, H. S., Kang, J. H., Hong, E. H., Kang, M. H., Seong, M., Cho, H., & Shin, Y. U. (2024) · Journal of Ophthalmology · doi:10.1155/2024/3654690 · Cited by 1 · see citing works
“ARPE-19 cells were obtained from the American Type Culture Collection and cultured in Dulbecco’s modifed Eagle’s medium/F-12 (DMEM/F12, Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS, Gibco) and 1% penicillin/streptomycin (Gibco) [13, 14].”
Method
ARP-T1 is a ciliogenesis protein associated with a novel ciliopathy in inherited basal cell cancer, Bazex-Dupré-Christol SyndromePreprint
Park, H., Papanastasi, E., Blanchard, G., Chiticariu, E., Bachmann, D., Plomann, M., Morice‐Picard, F., Vabres, P., Smahi, A., Huber, M., et al. (2019) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2019.12.16.877498

Induction of differentiation by pyruvate and DMEM in the human retinal pigment epithelium cell line ARPE-19
Ahmado, A., Carr, A.-J., Vugler, A. A., Semo, M., Gias, C., Lawrence, J. M., Chen, L. L., Chen, F. K., Turowski, P., da Cruz, L., et al. (2011) · Investigative Opthalmology & Visual Science · doi:10.1167/iovs.10-6374 · Cited by 100 · see citing works
Timeline of Achieving a Differentiated RPE Phenotype and Specialised Metabolism of ARPE-19 Cells Under Physiologically Relevant-Inducing Culture Conditions
Matei, I., Grimes, D., Suwanmanee, G., Bagnall, J., Spiller, D. G., & Paraoan, L. (2026) · Cells · doi:10.3390/cells15201846
Appropriately differentiated ARPE-19 cells regain phenotype and gene expression profiles similar to those of native RPE cells.
Samuel, W., Jaworski, C. J., Postnikova, O. A., Kutty, R. K., Duncan, T. E., Tan, L. X., Poliakov, E., Lakkaraju, A., & Redmond, T. M. (2017) · PubMed Central · Cited by 153 · see citing works
A Rapid protocol for the differentiation of human ARPE-19 cells
Hazim, R. A., Volland, S., & Williams, D. S. (2018) · Investigative Ophthalmology & Visual Science · Cited by 1 · see citing works
Expedited RPE‐derivation from human embryonic stem cells by partial PARP‐1 inhibition (LB55)
Pennington, B. O., & Clegg, D. (2014) · The FASEB Journal · doi:10.1096/fasebj.28.1_supplement.lb55

Authors: Hazim, R. A., Volland, S., Yen, A., Burgess, B. L., & Williams, D. S.
Affiliations: University of California, Los Angeles (US)
Corresponding author: Williams, David S.
ORCID: Hazim, Roni A.
Burgess, Barry L.
Williams, David S.
Venue: Experimental Eye Research
Publisher: Elsevier BV
Published: 2019-02
Volume: 179
Pages: 18-24
ISSN: 0014-4835 (ISSN-L)
DOI: 10.1016/j.exer.2018.10.009
PMC: PMC6360117
OpenAlex: W2897486375
Cited by: 126
References: 41
Citations by year: 2026: 16 · 2025: 24 · 2024: 26 · 2023: 16 · 2022: 15 · 2021: 7 · 2020: 18 · 2019: 4
FWCI: 7.29
Type: journal article
Language: en
Open access: green
www.ncbi.nlm.nih.gov
Licence: www.elsevier.com/tdm/userlicense/1.0/ (tdm)
www.elsevier.com/legal/tdmrep-license (tdm)
Locations: Experimental Eye Research (publishedVersion)
PubMed (publishedVersion)
PubMed Central (acceptedVersion)
eScholarship (California Digital Library) (submittedVersion)
Funders: National Institutes of Health (P30EY0331, R01EY013408, R01EY027442, F31EY026805)
Topics: Health Sciences › Medicine › Ophthalmology › Retinal Diseases and Treatments
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Retinal Development and Disorders
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Connexins and lens biology
Keywords: retinal pigment epithelium, RPE differentiation, nicotinamide, epithelial morphology, cytoskeletal organization, vitamin B metabolism
MeSH terms: Bestrophins, Retinoid Isomerohydrolase, Cell Differentiation, Cell Line, Cytoskeleton, Gene Expression Regulation, Humans, Microvilli, Niacinamide, Vitamin B Complex, Biomarkers, Fluorescent Antibody Technique, Indirect, cis-trans-Isomerases, Integrin beta Chains, Retinal Pigment Epithelium, Real-Time Polymerase Chain Reaction, Occludin, Zonula Occludens-1 Protein

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review

Rapid differentiation of the human RPE cell line, ARPE-19, induced by nicotinamide

Abstract

Integrity signals

Claims the paper makes

Claims other papers make about this one

Self-check

Cites

Cited by

Related

Record

About these checks