Validated and reproducible protocol for culturing single-eye porcine retinal pigment epithelium cells
Abstract
The study of retinal diseases and the development of treatments for conditions such as age-related macular degeneration (AMD) requires reliable and reproducible cell culture models. Porcine retinal pigment epithelium (RPE) cells have been widely used due to their similarities to human RPE cells, making them an ideal model for retinal studies. However, developing a standardized and reproducible protocol for culturing these cells remains a challenge. This article presents a validated protocol for cultivating single-eye porcine RPE cell cultures, ensuring consistency and reproducibility across experiments. The protocol addresses key considerations, including tissue isolation, cell maintenance, and characterization of the cultured cells, providing a useful resource for researchers in retinal biology and ophthalmology.
Keywords
Porcine RPE cells, retinal pigment epithelium, cell cultureHow to Cite
References
Wong, W.L.; Su, X.; Li, X.; Cheung, C.M.G.; Klein, R.; Cheng, C.-Y.; Wong, T.Y. Global prevalence of age-related macular degeneration and disease burden projection for 2020 and 2040: A systematic review and meta-analysis. Lancet Glob. Health 2014, 2, e106–e116. [Google Scholar] [CrossRef] [PubMed]
Zając-Pytrus, H.M.; Pilecka, A.; Turno-Kręcicka, A.; Adamiec-Mroczek, J.; Misiuk-Hojło, M. The Dry Form of Age-Related Macular Degeneration (AMD): The Current Concepts of Pathogenesis and Prospects for Treatment. Adv. Clin. Exp. Med. 2015, 24, 1099–1104. [Google Scholar] [CrossRef] [PubMed]
Fleckenstein, M.; Keenan, T.D.L.; Guymer, R.H.; Chakravarthy, U.; Schmitz-Valckenberg, S.; Klaver, C.C.; Wong, W.T.; Chew, E.Y. Age-related macular degeneration. Nat. Rev. Dis. Primers 2021, 7, 31. [Google Scholar] [CrossRef]
Bhutto, I.; Lutty, G. Understanding age-related macular degeneration (AMD): Relationships between the photoreceptor/retinal pigment epithelium/Bruch’s membrane/choriocapillaris complex. Mol. Asp. Med. 2012, 33, 295–317. [Google Scholar] [CrossRef]
Strauss, O. The retinal pigment epithelium in visual function. Physiol. Rev. 2005, 85, 845–881. [Google Scholar] [CrossRef]
Aisenbrey, S.; Zhang, M.; Bacher, D.; Yee, J.; Brunken, W.J.; Hunter, D.D. Retinal pigment epithelial cells synthesize laminins, including laminin 5, and adhere to them through alpha3-and alpha6-containing integrins. Investig. Ophthalmol. Vis. Sci. 2006, 47, 5537–5544. [Google Scholar] [CrossRef]
Schnichels, S.; Paquet-Durand, F.; Löscher, M.; Tsai, T.; Hurst, J.; Joachim, S.C.; Klettner, A. Retina in a dish: Cell cultures, retinal explants and animal models for common diseases of the retina. Prog. Retin. Eye Res. 2021, 81, 100880. [Google Scholar] [CrossRef]
Bharti, K.; den Hollander, A.I.; Lakkaraju, A.; Sinha, D.; Williams, D.S.; Finnemann, S.C.; Bowes-Rickman, C.; Malek, G.; D’Amore, P.A. Cell culture models to study retinal pigment epithelium-related pathogenesis in age-related macular degeneration. Exp. Eye Res. 2022, 222, 109170. [Google Scholar] [CrossRef]
Alge, C.S.; Hauck, S.M.; Priglinger, S.G.; Kampik, A.; Ueffing, M. Differential protein profiling of primary versus immortalized human RPE cells identifies expression patterns associated with cytoskeletal remodeling and cell survival. J. Proteome Res. 2006, 5, 862–878. [Google Scholar] [CrossRef]
Dunn, K.C.; Aotaki-Keen, A.E.; Putkey, F.R.; Hjelmeland, L.M. ARPE-19, a human retinal pigment epithelial cell line with differentiated properties. Exp. Eye Res. 1996, 62, 155–169. [Google Scholar] [CrossRef]
Tian, J.; Ishibashi, K.; Honda, S.; Boylan, S.A.; Hjelmeland, L.M.; Handa, J.T. The expression of native and cultured human retinal pigment epithelial cells grown in different culture conditions. Br. J. Ophthalmol. 2005, 89, 1510–1517. [Google Scholar] [CrossRef] [PubMed]
Ahmado, A.; Carr, A.-J.; Vugler, A.A.; Gias, C.; Lawrence, J.M.; Chen, L.L.; Chen, F.K.; Turowski, P.; da Cruz, L.; Coffey, P.J. Induction of differentiation by pyruvate and DMEM in the human retinal pigment epithelium cell line ARPE-19. Investig. Ophthalmol. Vis. Sci. 2011, 52, 7148–7159. [Google Scholar] [CrossRef] [PubMed]
Samuel, W.; Jaworski, C.; Postnikova, O.A.; Kutty, R.K.; Duncan, T.; Tan, L.X.; Poliakov, E.; Lakkaraju, A.; Redmond, T.M. Appropriately differentiated ARPE-19 cells regain phenotype and gene expression profiles similar to those of native RPE cells. Mol. Vis. 2017, 23, 60–89. [Google Scholar] [PubMed]
Golconda, P.; Andrade-Medina, M.; Oberstein, A. Subconfluent ARPE-19 Cells Display Mesenchymal Cell-State Characteristics and Behave like Fibroblasts, Rather Than Epithelial Cells, in Experimental HCMV Infection Studies. Viruses 2023, 16, 49. [Google Scholar] [CrossRef]
Luo, Y.; Zhuo, Y.; Fukuhara, M.; Rizzolo, L.J. Effects of culture conditions on heterogeneity and the apical junctional complex of the ARPE-19 cell line. Investig. Ophthalmol. Vis. Sci. 2006, 47, 3644–3655. [Google Scholar] [CrossRef]
Nakai-Futatsugi, Y.; Jin, J.; Ogawa, T.; Sakai, N.; Maeda, A.; Hironaka, K.-I.; Fukuda, M.; Danno, H.; Tanaka, Y.; Hori, S.; et al. Pigmentation level of human iPSC-derived RPE does not indicate a specific gene expression profile. eLife 2024, 12, RP92510. [Google Scholar] [CrossRef]
Farjood, F.; Manos, J.D.; Wang, Y.; Williams, A.L.; Zhao, C.; Borden, S.; Alam, N.; Prusky, G.; Temple, S.; Stern, J.H.; et al. Identifying biomarkers of heterogeneity and transplantation efficacy in retinal pigment epithelial cells. J. Exp. Med. 2023, 220, e20230913. [Google Scholar] [CrossRef]
Miyagishima, K.J.; Wan, Q.; Corneo, B.; Sharma, R.; Lotfi, M.R.; Boles, N.C.; Hua, F.; Maminishkis, A.; Zhang, C.; Blenkinsop, T.; et al. In Pursuit of Authenticity: Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium for Clinical Applications. Stem Cells Transl. Med. 2016, 5, 1562–1574. [Google Scholar] [CrossRef]
Rzhanova, L.A.; Markitantova, Y.V.; Aleksandrova, M.A. Recent Achievements in the Heterogeneity of Mammalian and Human Retinal Pigment Epithelium: In Search of a Stem Cell. Cells 2024, 13, 281. [Google Scholar] [CrossRef]
Luo, M.; Chen, Y. Application of stem cell-derived retinal pigmented epithelium in retinal degenerative diseases: Present and future. Int. J. Ophthalmol. 2018, 11, 150–159. [Google Scholar]
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