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The Retinal Pigment Epithelial Cell Line (ARPE-19) Displays Mosaic Structural Chromosomal Aberrations

  • Elizaveta Fasler-Kan
  • Nijas Aliu
  • Kerstin Wunderlich
  • Sylvia Ketterer
  • Sabrina Ruggiero
  • Steffen Berger
  • Peter Meyer
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1745)

Abstract

The retinal pigment epithelial cell line ARPE-19 was established in 1996 and remains widely used today for biomedical and in particular ophthalmology research. We have analyzed the chromosomes of the ARPE-19 cell line and found cultured cells exist as a heterogeneous mixture having both normal karyotypes and chromosomal rearrangements. In ARPE-19 cells, we observed metaphases with a single translocation t(15;19) and metaphases with two translocations t(5;15) and t(15;19) and a derivative chromosome 9. Aneuploidies have also been detected (monosomy: −16; trisomy: +11, +18). Multiple attempts to isolate clones with a normal karyotype from those with aberrant karyotypes failed due to senescence of cells of normal karyotypes. We could, however, isolate clones with the translocation t(15;19) and clones with two translocations t(5;15) and t(15;19). In continued cell culture after second subcloning for 30 passages, all clones maintained their cytogenetic integrity.

We have further investigated the chromosomal profiles of the ARPE-19 cell line from another laboratory and observed cells with a normal karyotype as well as abnormalities in chromosomes 6p and 11q. The DNA profiles of the ARPE-19 cells from both labs were identical to the ATCC profiles, excluding contamination with other cell lines. Since chromosomal translocations in ARPE-19 cells differ from lab to lab and display a mosaicism for structural chromosomal aberrations, researchers dealing with ARPE-19 cells should screen their stocks for chromosomal aberrations and proceed with caution against misinterpretations during experimental manipulations with this cell line. This chapter describes in detail our laboratory methods for single cell cloning, karyotype analysis and fluorescence in situ hybridization (FISH), which we used for the identification and characterization of chromosomal translocations in the retinal pigment epithelial cell line ARPE-19.

Keywords

Retinal pigment cells ARPE-19 cell line Karyotype Chromosomal translocations Chromosomal aberrations FISH Single-cell cloning Heterogeneity Mosaicism 

Notes

Acknowledgments

The work was supported in part by Grant-in-Aid (University of Bern) and Batzebär grant. We are thankful to Friedel Wenzel (Cytogenetics Department, University of Basel) for his help. We are also grateful to Dr. Ron Tynes (University of Applied Sciences Northwestern Switzerland) for his help with the editing and preparation of the manuscript.

References

  1. 1.
    Pascolini D, Mariotti SP (2012) Global estimates of visual impairment:2010. Br J Ophthalmol 96(5):614–618.  https://doi.org/10.1136/bjophthalmol-2011-300539.
  2. 2.
    Alfaro DV, Liggett PE, Mieler WF, Quiroz-Mercado H, Jager RD, Tano Y (eds) (2006) Age-related macular degeneration: a comprehensive textbook. Lippincott Williams & Wilkins, PhiladelphiaGoogle Scholar
  3. 3.
    Klein R, Klein BE, Linton KL (1992) Prevalence of age-related maculopathy. The Beaver Dam Eye Study. Ophthalmology 99(6):933–943CrossRefPubMedGoogle Scholar
  4. 4.
    Ding X, Patel M, Chan CC (2009) Molecular pathology of age-related macular degeneration. Prog Retin Eye Res 28(1):1–18.  https://doi.org/10.1016/j.preteyeres.2008.10.001 CrossRefPubMedGoogle Scholar
  5. 5.
    van Leewen R, Klaver CC, Vingerling JR, Hofman A, de Jong PT (2003) Epidemiology of age-related maculopathy: a review. Eur J Epidemiol 18(9):845–854CrossRefGoogle Scholar
  6. 6.
    Hamdi HK, Kenney C (2003) Age-related macular degeneration: a new viewpoint. Front Biosci 8:e305–e314CrossRefPubMedGoogle Scholar
  7. 7.
    Jager RD, Mieler WF, Miller JW (2008) Age-related macular degeneration. N Engl J Med 358:2606–2617.  https://doi.org/10.1056/NEJMra0801537 CrossRefPubMedGoogle Scholar
  8. 8.
    Dunn KC, Aotaki-Keen AE, Putkey FR, Hjelmeland LM (1996) A human retinal pigment epithelial cell line with differentiated properties. Exp Eye Res 62:155–169CrossRefPubMedGoogle Scholar
  9. 9.
    Dunn KC, Marmorstein AD, Bonilha VL, Rodriguez-Boulan E, Giordano F, Hjelmeland LM (1998) Use of the ARPE-19 cell line as a model of RPE polarity: basolateral secretion of FGF5. Invest Ophthalmol Vis Sci 39:2744–2749PubMedGoogle Scholar
  10. 10.
    Stepanenko AA, Dmitrenko VV (2015) HEK-293 in cell biology and cancer research: phenotype, karyotype, tumorigenicity, and stress-induced genome-phenotype evolution. Gene 569:182–190.  https://doi.org/10.1016/j.gene.2015.05.065 CrossRefPubMedGoogle Scholar
  11. 11.
    Shen C, Gu M, Song C, Miao L, Hu L, Liang D, Zheng C (2008) The tumorigenicity diversification in human embryonic kidney cell line cultured in vitro. Biologicals 36(4):263–268.  https://doi.org/10.1016/j.biologicals.2008.02.002 CrossRefPubMedGoogle Scholar
  12. 12.
    Ritter A, Voedisch B, Wienberg J, Wilms B, Geisse S, Jostock T, Laux H (2016) Deletion of a telomeric region on chromosome 8 correlates with higher productivity and stability of CHO cell lines. Biotechnol Bioeng 113(5):1084–1093.  https://doi.org/10.1002/bit.25876 CrossRefPubMedGoogle Scholar
  13. 13.
    Diaferia GR, Conti L, Redaelli S, Cattaneo M, Mutti C, DeBlasio P, Dalpra L, Cattaneo E, Biunno I (2011) Systematic chromosomal analysis of cultured mouse neural stem cell lines. Stem Cells Dev 20(8):1411–1423.  https://doi.org/10.1089/scd.2010.0359 CrossRefPubMedGoogle Scholar
  14. 14.
    Harrison N (2012) Genetic instability in neural stem cells: an inconvenient truth? J Clin Invest 122(2):484–486.  https://doi.org/10.1172/JCI62002 CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  • Elizaveta Fasler-Kan
    • 1
    • 2
  • Nijas Aliu
    • 3
  • Kerstin Wunderlich
    • 1
    • 4
  • Sylvia Ketterer
    • 1
  • Sabrina Ruggiero
    • 2
  • Steffen Berger
    • 2
  • Peter Meyer
    • 4
  1. 1.Department of BiomedicineUniversity of Basel and University Hospital BaselBaselSwitzerland
  2. 2.Department of Pediatric SurgeryInselspital, University Hospital Bern and Department of Biomedical Research, University of BernBernSwitzerland
  3. 3.Department of Human GeneticsUniversity Children’s Hospital, InselspitalBernSwitzerland
  4. 4.Department of OphthalmologyUniversity of BaselBaselSwitzerland

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