دورية أكاديمية
أعرض في EDS

A convenient protocol for establishing a human cell culture model of the outer retina.

التفاصيل البيبلوغرافية
العنوان: A convenient protocol for establishing a human cell culture model of the outer retina.
المؤلفون: Lynn SA; Clinical and Experimental Sciences, Faculty of Medicine, MP 806, Tremona Road, University of Southampton, Southampton, Hampshire, SO16 6YD, UK., Keeling E; Clinical and Experimental Sciences, Faculty of Medicine, MP 806, Tremona Road, University of Southampton, Southampton, Hampshire, SO16 6YD, UK., Dewing JM; Clinical and Experimental Sciences, Faculty of Medicine, MP 806, Tremona Road, University of Southampton, Southampton, Hampshire, SO16 6YD, UK., Johnston DA; Biomedical Imaging Unit, MP 806, Tremona Road, University of Southampton, Southampton, Hampshire, SO16 6YD, UK., Page A; Biomedical Imaging Unit, MP 806, Tremona Road, University of Southampton, Southampton, Hampshire, SO16 6YD, UK., Cree AJ; Clinical and Experimental Sciences, Faculty of Medicine, MP 806, Tremona Road, University of Southampton, Southampton, Hampshire, SO16 6YD, UK., Tumbarello DA; Biological Sciences, Faculty of Natural & Environmental Sciences, Life Sciences Building 85, University of Southampton, Southampton, Hampshire, SO17 1BJ, UK., Newman TA; Clinical and Experimental Sciences, Faculty of Medicine, MP 806, Tremona Road, University of Southampton, Southampton, Hampshire, SO16 6YD, UK., Lotery AJ; Clinical and Experimental Sciences, Faculty of Medicine, MP 806, Tremona Road, University of Southampton, Southampton, Hampshire, SO16 6YD, UK.; Eye Unit, University Hospital Southampton NHS Foundation Trust, Southampton, Hampshire, SO16 6YD, UK., Ratnayaka JA; Clinical and Experimental Sciences, Faculty of Medicine, MP 806, Tremona Road, University of Southampton, Southampton, Hampshire, SO16 6YD, UK.
المصدر: F1000Research [F1000Res] 2018 Jul 18; Vol. 7, pp. 1107. Date of Electronic Publication: 2018 Jul 18 (Print Publication: 2018).
نوع المنشور: Journal Article; Research Support, Non-U.S. Gov't
اللغة: English
بيانات الدورية: Publisher: F1000 Research Ltd Country of Publication: England NLM ID: 101594320 Publication Model: eCollection Cited Medium: Internet ISSN: 2046-1402 (Electronic) Linking ISSN: 20461402 NLM ISO Abbreviation: F1000Res Subsets: MEDLINE
أسماء مطبوعة: Original Publication: London : F1000 Research Ltd
مواضيع طبية MeSH: Extracellular Matrix*/metabolism , Extracellular Matrix*/pathology , Models, Biological* , Retinal Pigment Epithelium*/metabolism , Retinal Pigment Epithelium*/pathology, Macular Degeneration/*metabolism, Animals ; Bruch Membrane/metabolism ; Bruch Membrane/pathology ; Cell Adhesion ; Cell Culture Techniques/methods ; Cell Line ; Humans ; Macular Degeneration/pathology ; Swine
مستخلص: The retinal pigment epithelium (RPE) plays a key role in the pathogenesis of several blinding retinopathies. Alterations to RPE structure and function are reported in Age-related Macular Degeneration, Stargardt and Best disease as well as pattern dystrophies. However, the precise role of RPE cells in disease aetiology remains incompletely understood. Many studies into RPE pathobiology have utilised animal models, which only recapitulate limited disease features. Some studies are also difficult to carry out in animals as the ocular space remains largely inaccessible to powerful microscopes. In contrast, in-vitro models provide an attractive alternative to investigating pathogenic RPE changes associated with age and disease. In this article we describe the step-by-step approach required to establish an experimentally versatile in-vitro culture model of the outer retina incorporating the RPE monolayer and supportive Bruch's membrane (BrM). We show that confluent monolayers of the spontaneously arisen human ARPE-19 cell-line cultured under optimal conditions reproduce key features of native RPE. These models can be used to study dynamic, intracellular and extracellular pathogenic changes using the latest developments in microscopy and imaging technology. We also discuss how RPE cells from human foetal and stem-cell derived sources can be incorporated alongside sophisticated BrM substitutes to replicate the aged/diseased outer retina in a dish. The work presented here will enable users to rapidly establish a realistic in-vitro model of the outer retina that is amenable to a high degree of experimental manipulation which will also serve as an attractive alternative to using animals. This in-vitro model therefore has the benefit of achieving the 3Rs objective of reducing and replacing the use of animals in research. As well as recapitulating salient structural and physiological features of native RPE, other advantages of this model include its simplicity, rapid set-up time and unlimited scope for detailed single-cell resolution and matrix studies.
Competing Interests: No competing interests were disclosed.
References: J Cell Sci. 1993 Jan;104 ( Pt 1):37-49. (PMID: 8383696)
Invest Ophthalmol Vis Sci. 2004 May;45(5):1281-8. (PMID: 15111578)
Invest Ophthalmol Vis Sci. 2011 Sep 09;52(10):7148-59. (PMID: 21743014)
J Biomed Mater Res A. 2010 Dec 15;95(4):1233-43. (PMID: 20939049)
Cytometry A. 2015 Jun;87(6):568-79. (PMID: 25605428)
J Ophthalmol. 2014;2014:801787. (PMID: 25177495)
Am J Pathol. 1998 Nov;153(5):1641-6. (PMID: 9811357)
Proc Natl Acad Sci U S A. 2005 Aug 16;102(33):11900-5. (PMID: 16079201)
Adv Exp Med Biol. 2012;724:15-36. (PMID: 22411231)
Exp Eye Res. 2014 Jul;124:74-85. (PMID: 24861273)
Prog Retin Eye Res. 2011 Sep;30(5):296-323. (PMID: 21704180)
Eur J Pharm Sci. 2010 Jul 11;40(4):289-96. (PMID: 20385230)
Nat Protoc. 2016 Jul;11(7):1206-18. (PMID: 27281648)
Cells. 2018 Feb 23;7(2):. (PMID: 29473871)
Sci Rep. 2017 Jul 7;7(1):4901. (PMID: 28687758)
Invest Ophthalmol Vis Sci. 2010 Jan;51(1):553-60. (PMID: 19696182)
Exp Eye Res. 2014 Sep;126:5-15. (PMID: 25152359)
Br J Ophthalmol. 2011 Apr;95(4):563-8. (PMID: 19965827)
Proc Natl Acad Sci U S A. 2015 Jun 9;112(23):E3040-9. (PMID: 25991857)
Exp Eye Res. 2014 Sep;126:1-4. (PMID: 25152358)
Prog Retin Eye Res. 2010 May;29(3):169-90. (PMID: 20206286)
Proc Natl Acad Sci U S A. 2011 Nov 8;108(45):18277-82. (PMID: 21969589)
Mol Vis. 2017 Mar 05;23:60-89. (PMID: 28356702)
Invest Ophthalmol Vis Sci. 2006 Aug;47(8):3612-24. (PMID: 16877436)
J Exp Med. 2001 Nov 5;194(9):1289-98. (PMID: 11696594)
Exp Eye Res. 1996 Feb;62(2):155-69. (PMID: 8698076)
Biophys J. 2004 Jun;86(6):3993-4003. (PMID: 15189895)
Mol Vis. 2009;15:283-95. (PMID: 19204785)
Tissue Cell. 2017 Aug;49(4):447-460. (PMID: 28669519)
Methods Mol Biol. 2013;935:285-95. (PMID: 23150376)
Exp Eye Res. 2017 Dec;165:35-46. (PMID: 28847738)
Methods Mol Biol. 2013;945:45-65. (PMID: 23097100)
Invest Ophthalmol Vis Sci. 2001 Jul;42(8):1685-90. (PMID: 11431429)
Exp Eye Res. 2014 Sep;126:51-60. (PMID: 24780752)
Invest Ophthalmol Vis Sci. 2014 Feb 14;55(2):926-34. (PMID: 24458156)
Proc Natl Acad Sci U S A. 1996 Oct 1;93(20):10779-84. (PMID: 8855257)
J Biomed Biotechnol. 2010;2010:402593. (PMID: 20224657)
Nat Commun. 2017 May 19;8:15374. (PMID: 28524846)
Mol Aspects Med. 2012 Aug;33(4):487-509. (PMID: 22705444)
Invest Ophthalmol Vis Sci. 2011 Nov 04;52(12):8614-20. (PMID: 21960553)
Am J Physiol Cell Physiol. 2011 Apr;300(4):C723-42. (PMID: 21209361)
PLoS One. 2015 Apr 29;10(4):e0125631. (PMID: 25923208)
Mol Vis. 2011;17:1701-15. (PMID: 21738400)
Sci Rep. 2018 Apr 13;8(1):5944. (PMID: 29654292)
Proc Natl Acad Sci U S A. 2017 Sep 26;114(39):E8214-E8223. (PMID: 28878022)
Clin Ophthalmol. 2008 Jun;2(2):413-24. (PMID: 19668732)
Mol Aspects Med. 2012 Aug;33(4):295-317. (PMID: 22542780)
Acta Histochem Cytochem. 2007 Aug 30;40(4):101-11. (PMID: 17898874)
Exp Eye Res. 1987 Dec;45(6):907-22. (PMID: 2828096)
Mol Vis. 2009;15:223-34. (PMID: 19180257)
J Cell Mol Med. 2013 Jul;17(7):833-43. (PMID: 23663427)
Proc Natl Acad Sci U S A. 1997 Nov 25;94(24):12932-7. (PMID: 9371778)
Nat Protoc. 2009;4(5):662-73. (PMID: 19373231)
Invest Ophthalmol Vis Sci. 2014 May 20;55(6):3709-19. (PMID: 24845632)
Mol Vis. 2001 Feb 07;7:14-9. (PMID: 11182021)
Exp Eye Res. 2014 Sep;126:16-26. (PMID: 24731966)
Biochem Pharmacol. 2018 May;151:144-156. (PMID: 29309763)
F1000Res. 2018 Jul 18;7:1107. (PMID: 30271583)
معلومات مُعتمدة: NC/L001152/1 United Kingdom NC3RS_ National Centre for the Replacement, Refinement and Reduction of Animals in Research
فهرسة مساهمة: Keywords: Cell culture; Disease modelling; In-vitro; Retinal Pigment Epithelium (RPE); Retinopathy
تواريخ الأحداث: Date Created: 20181002 Date Completed: 20190904 Latest Revision: 20230928
رمز التحديث: 20230928
مُعرف محوري في PubMed: PMC6137423
DOI: 10.12688/f1000research.15409.1
PMID: 30271583
قاعدة البيانات: MEDLINE
الوصف
تدمد:2046-1402
DOI:10.12688/f1000research.15409.1