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M. W. Djojosubroto, M. Tekaya, D. Wanner, F. Bollotte, P. Wirapati, F. Radtke, Y. Arsenijevic; Increased Chromosomal Aberrations and Transformation of Adult Mouse Retinal Stem Cells. Invest. Ophthalmol. Vis. Sci. 2008;49(13):3548.
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© ARVO (1962-2015); The Authors (2016-present)
The potential of stem cell therapy for degenerative diseases is highly recognized. Generation of high numbers of retinal stem cells (RSCs) in vitro would thus be beneficial for retina transplantation. Yet, cells in cultivation can be unstable and have a risk of transformation. We analyzed RSCs isolated from neonatal retinas and pigmented ciliary margin of adult mouse eyes.
Neonatal and adult RSCs were isolated and cultured as we previously reported. Chromosomal counting and painting were done to examine chromosomal aberrations along cell passaging. Analyses of anchorage-independent cell proliferation in soft agarose assay and telomerase activity (TRAP Assay) were performed to evaluate cell transformation potentials. Three cell lines were subcutaneously transplanted into nude mice and the histology of tumors were assessed.
Both neonatal and adult RSCs are highly proliferative. Most neonatal RSCs (83%) at passage 9 have 40 chromosomes, yet only 73% have normal karyotype by chromosomal painting analysis. At passage 36, 33% retain normal chromosomal number, however all cells have translocation of chromosome 16 to 9. Surprisingly, adult mouse RSCs show more chromosomal aberrations even at early passage. The chromosome number highly varies, with a large proportion of aneuploidy. All cell lines show telomerase activity and anchorage-dependent cell proliferation, albeit to different levels. Adult RSCs formed tumors within 4-8 weeks, but no tumor was formed by neonatal RSCs up to 6 months. Interestingly, our data indicates a preference for (pseudo)diploidy vs. polyploidy in latepassaged adult transformed cells. Moreover, we found indication of chromosome missegregation and formation of anaphase bridges during mitosis.
Our data indicate that neonatal and adult RSCs have different levels of chromosomal stability, and adult RSCs may have higher chance of spontaneous transformation in vitro.
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