Retinal Cell Line (R28)

The R28 cells were developed from E1A-NR.3 parental cell line through three rounds of limiting dilution and were therefore derived from a single cell. Despite their clonal origin, these cells display both glial and neuronal cell markers indicative of a retinal precursor cell.

The parental line E1A-NR.3 was established by immortalization of postnatal day 6 rat neuroretinal tissue using the psi2 replication incompetent retroviral vector. As a result, these cells are already resistant to geneticin/G418 and would require an alternative selection marker for transfection studies. These cells were designed not to form tumors in animals.

From the laboratory of Gail M. Seigel, PhD, University of Rochester.

You can now pre-order Dr. Seigel's book Academania: My Life in the Trenches of Biomedical Research.


New publication: R28 Retinal Precursor Cells: The First 20 Years

New publication: A Microarray Dataset of Genes Expressed by the R28 Retinal Cell Line

Read Dr. Seigel's related blog post, On the Road to Retinal Cells.

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Retinal Cell Line (R28)
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Product Type: Cell Line
Name: R28
Cell Type: 12S E1A-immmortalized rat retinal cells
Accession ID: CVCL_5I35
Source: Postnatal day 6 rat neural retina
Organism: Rat; NOTE: The R28 cells have undergone verification by IDEXX-RADIL to be of rat origin without contamination by other mammalian cell lines. A baseline genetic profile of these cells is available upon request.
Morphology: Adherent with glial and neuronal morphologies
Biosafety Level: BSL-1
Subculturing: Split 1:2 when 80% confluent (See also: R28 Cell Care Instructions)
Growth Conditions: DMEM+; See recipe in the recommended SOP:

  R28 Cell Care Instructions

Cryopreservation: 90% culture medium with 10% DMSO
Storage: -150C
Shipped: Dry ice

From the laboratory of Gail M. Seigel, PhD, University of Rochester.

R28-immortalized cell characterization

  1. Seigel, GM, 1996. Establishment of an E1A-immortalized rat retinal cell culture, In Vitro Cell. Devel. Biol. 32: 66-68.
  2. Seigel, GM, Mutchler, A.L., and E.L. Imperato.  1996. Expression of glial cell markers in a retinal precursor cell line. Mol. Vis. 2: 2.
  3. Seigel, GM, Takahashi, M., Adamus, G., McDaniel, T. 1998.  Intraocular transplantation of E1A-immortalized retinal precursor cells. Cell Transplant. 7 (6):  559-566.
  4. Seigel, GM, Sun, W, Wang, J., Hershberger, DH, Campbell, LM, Salvi, RJ. 2004. Neuronal gene expression and function in the growth-stimulated R28 retinal precursor cell line.  Curr. Eye Res., 28 (4):257-269.
  5. Seigel, GM and Salvi, RJ. A microarray dataset of genes expressed by the R28 retinal precursor cell line. Dataset Papers in Neuroscience, 2013, ID: 261063. View Article
  6. Seigel GM. R28 retinal precursor cells: The first 20 years. Mol Vis. 2014; 20: 301-306. View Article

R28-immortalized cell utilization

  1. Uddin MI, Evans SM, Craft JR, Capozzi ME, McCollum GW, Yang R, Marnett LJ,Uddin MJ, Jayagopal A, Penn JS. In Vivo Imaging of Retinal Hypoxia in a Model of Oxygen-Induced Retinopathy. Sci Rep. 2016 Aug 5;6:31011. View Article
  2. Su PJ, Liu Z, Zhang K, Han X, Saito Y, Xia X, Yokoi K, Shen H, Qin L. Retinal synaptic regeneration via microfluidic guiding channels. Sci Rep. 2015 Aug28;5:13591. doi: 10.1038/srep13591. View Article
  3. Song Y, Hong S, Iizuka Y, Kim CY, Seong GJ. The neuroprotective effect of maltol against oxidative stress on rat retinal neuronal cells. Korean J Ophthalmol. 2015 Feb;29(1):58-65. doi: 10.3341/kjo.2015.29.1.58. View Article
  4. Identification of pigment epithelium-derived factor protein forms with distinct activities on tumor cell lines. Subramanian P, Deshpande M, Locatelli-Hoops S, Moghaddam-Taaheri S, Gutierrez D, Fitzgerald DP, Guerrier S, Rapp M, Notario V, Becerra SP. J Biomed Biotechnol. 2012;2012:425907. Epub 2012 Jun 4.
  5. Mouse acetylcholinesterase enhances neurite outgrowth of rat R28 cells through interaction with laminin-1.  Sperling LE, Klaczinski J, Schütz C, Rudolph L, Layer PG. PLoS One. 2012;7(5):e36683. Epub 2012 May 3.
  6. Blue light stress in retinal neuronal (R28) cells is dependent on wavelength range and irradiance.  Knels L, Valtink M, Roehlecke C, Lupp A, de la Vega J, Mehner M, Funk RH. Eur J Neurosci. 2011 Aug;34(4):548-58. doi: 10.1111/j.1460-9568.2011.07790.x. Epub 2011 Jul 22.
  7. Inhibition of reactive gliosis prevents neovascular growth in the mouse model of oxygen-induced retinopathy. DeNiro M, Al-Mohanna FH, Al-Mohanna FA.  PLoS One. 2011;6(7):e22244. Epub 2011 Jul 14.
  8. The effects of commercially available preservative-free FDA-approved triamcinolone (Triesence®) on retinal cells in culture. Zacharias LC, Estrago-Franco MF, Ramirez C, Kenney MC, Takahashi WY, Seigel GM, Kuppermann BD.  J Ocul Pharmacol Ther. 2011 Apr;27(2):143-50.
  9. Boriushkin E, Wang JJ, Li J, Jing G, Seigel GM, Zhang SX. Identification of p58IPK as a novel neuroprotective factor for retinal neurons. Invest Ophthalmol Vis Sci. 2015 Feb 5;56(2):1374-86. View Article
  10. Chan JF, Yip CC, Tsang JO, Tee KM, Cai JP, Chik KK, Zhu Z, Chan CC, Choi GK, Sridhar S, Zhang AJ, Lu G, Chiu K, Lo AC, Tsao SW, Kok KH, Jin DY, Chan KH, Yuen KY. Differential cell line susceptibility to the emerging Zika virus: implications for disease pathogenesis, non-vector-borne human transmission and animal reservoirs. Emerg Microbes Infect. 2016 Aug 24;5:e93. View Article
  11. Evans SM, Kim K, Moore CE, Uddin MI, Capozzi ME, Craft JR, Sulikowski GA, Jayagopal A. Molecular probes for imaging of hypoxia in the retina. Bioconjug Chem. 2014 Nov 19;25(11):2030-7. doi: 10.1021/bc500400z. PubMed PMID: 25250692; PubMed Central PMCID: PMC4240343. View Article
  12. Kenealey J, Subramanian P, Comitato A, Bullock J, Keehan L, Polato F, Hoover D, Marigo V, Becerra SP. Small Retinoprotective Peptides Reveal a Receptor-binding Region on Pigment Epithelium-derived Factor. J Biol Chem. 2015 Oct 16;290(42):25241-53. View Article
  13. Uddin MI, Evans SM, Craft JR, Marnett LJ, Uddin MJ, Jayagopal A. Applications of azo-based probes for imaging retinal hypoxia. ACS Med Chem Lett. 2015 Feb 12;6(4):445-9. View Article
  14. Asnaghi L, Tripathy A, Yang Q, Kaur H, Hanaford A, Yu W, Eberhart CG. Targeting Notch signaling as a novel therapy for retinoblastoma. Oncotarget. 2016 Sep 20. View Article
  15. Taylor-Walker G, Lynn SA, Keeling E, Munday R, Johnston DA, Page A, Scott JA, Goverdhan S, Lotery AJ, Ratnayaka JA. The Alzheimer's-related amyloid beta peptide is internalised by R28 neuroretinal cells and disrupts the microtubule associated protein 2 (MAP-2). Exp Eye Res. 2016 Dec;153:110-121. View Article
  16. Winokur PN, Subramanian P, Bullock JL, Arocas V, Becerra SP. Comparison of two neurotrophic serpins reveals a small fragment with cell survival activity. Mol Vis. 2017 Jul 3;23:372-384. eCollection 2017. View Article
  17. Davis BM, Pahlitzsch M, Guo L, Balendra S, Shah P, Ravindran N, Malaguarnera G, Sisa C, Shamsher E, Hamze H, Noor A, Sornsute A, Somavarapu S, Cordeiro MF. Topical Curcumin Nanocarriers are Neuroprotective in Eye Disease. Sci Rep. 2018 Jul 23;8(1):11066. View Article
  18. Mathew B, Ravindran S, Liu X, Torres L, Chennakesavalu M, Huang CC, Feng L, Zelka R, Lopez J, Sharma M, Roth S. Mesenchymal stem cell-derived extracellular vesicles and retinal ischemia-reperfusion. Biomaterials. 2019 Jan 9;197:146-160. View Article
  19. Arranz-Romera A, Davis BM, Bravo-Osuna I, Esteban-Pérez S, Molina-Martínez IT, Shamsher E, Ravindran N, Guo L, Cordeiro MF, Herrero-Vanrell R. Simultaneous co-delivery of neuroprotective drugs from multi loaded PLGA microspheres for the treatment of glaucoma. J Control Release. 2019 Mar 10;297:26-38. View Article
  20. Núñez-Álvarez C, Osborne NN. Blue light exacerbates and red light counteracts negative insults to retinal ganglion cells in situ and R28 cells in vitro. Neurochem Int. 2019 May;125:187-196. View Article
  21. Chang C, Hu Y, Hogan SL, et al. Pharmacogenomic Variants May Influence the Urinary Excretion of Novel Kidney Injury Biomarkers in Patients Receiving Cisplatin. Int J Mol Sci. 2017;18(7):1333. Published 2017 Jun 22. View Article
  22. Bullock J, Pagan-Mercado G, Becerra SP. Cell-based assays to identify novel retinoprotective agents. MethodsX. 2020 Aug 10;7:101026. View Article
  23. Kim HM, Kwon J, Lee K, Lee JW, Jang DS, Kwon HC. Constituents of Gastrodia elata and Their Neuroprotective Effects in HT22 Hippocampal Neuronal, R28 Retinal Cells, and BV2 Microglial Cells. Plants (Basel). 2020 Aug 18;9(8):1051.  View article
  24. Yadav M, Schiavone N, Guzman-Aranguez A, Giansanti F, Papucci L, Perez de Lara MJ, Singh M, Kaur IP. Atorvastatin-loaded solid lipid nanoparticles as eye drops: proposed treatment option for age-related macular degeneration (AMD). Drug Deliv Transl Res. 2020 Aug;10(4):919-944. View article
  25. Yan B, Gao L, Huang Y, Wang X, Lang X, Yan F, Meng B, Sun X, Li G, Wang Y. Exosomes derived from BDNF-expressing 293T attenuate ischemic retinal injury in vitro and in vivo. Aging (Albany NY). 2020 Nov 29;12. View article 

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