Genetic barcoding reveals clonal dominance in iPSC-derived mesenchymal stromal cells.

Stem Cell Research & Therapy
Jonathan HollmannWolfgang Wagner

Abstract

The use of mesenchymal stromal cells (MSCs) for research and clinical application is hampered by cellular heterogeneity and replicative senescence. Generation of MSC-like cells from induced pluripotent stem cells (iPSCs) may circumvent these limitations, and such iPSC-derived MSCs (iMSCs) are already tested in clinical trials. So far, a comparison of MSCs and iMSCs was particularly addressed in bulk culture. Despite the high hopes in cellular therapy, only little is known how the composition of different subclones changes in these cell preparations during culture expansion. In this study, we used multicolor lentiviral genetic barcoding for the marking of individual cells within cell preparations. Based on this, we could track the clonal composition of syngenic MSCs, iPSCs, and iMSCs during culture expansion. Furthermore, we analyzed DNA methylation patterns at senescence-associated genomic regions by barcoded bisulfite amplicon sequencing. The proliferation and differentiation capacities of individual subclones within MSCs and iMSCs were investigated with limiting dilution assays. Overall, the clonal composition of primary MSCs and iPSCs gradually declined during expansion. In contrast, iMSCs became oligoclonal early during dif...Continue Reading

References

Mar 26, 2008·Molecular Therapy : the Journal of the American Society of Gene Therapy·Kristoffer WeberBoris Fehse
Jun 1, 2011·Cell Stem Cell·Chris E P GoldringB Kevin Park
Dec 14, 2011·Cytotherapy·Anne SchellenbergWolfgang Wagner
Jan 10, 2012·Aging Cell·Carmen M KochWolfgang Wagner
Apr 7, 2012·Nature Protocols·Kristoffer WeberBoris Fehse
Jun 30, 2012·Nature Methods·Johannes SchindelinAlbert Cardona
Oct 20, 2012·Genome Research·Carmen M KochWolfgang Wagner
Jan 31, 2014·Nucleic Acids Research·Kerstin CornilsBoris Fehse
Jun 30, 2014·Nature Methods·Aryeh WarmflashAli H Brivanlou
Dec 31, 2014·Proceedings of the National Academy of Sciences of the United States of America·Qingguo ZhaoFei Liu
Mar 11, 2015·Scientific Reports·Michael LenzWolfgang Wagner
Jun 20, 2018·Stem Cell Research & Therapy·David Brenière-LetuffeJustus Stenzig
Oct 8, 2018·Nature Reviews. Cancer·Zbyslaw SondkaSimon A Forbes
Jul 16, 2019·Stem Cell Reports·Anton SelichMichael Rothe
Jan 28, 2020·Stem Cell Reports·Eduardo Fernandez-RebolloWolfgang Wagner

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Methods Mentioned

BETA
fluorescence-activated cell sorter
flow cytometry
PCR
Amplicon Seq
fluorescence microscopy
amplicon sequencing

Software Mentioned

Pluri Score
custom
GraphPad Prism
ImageJ
Epi
cutadapt
Calc Limiting Dilution
R package vegan
ggplot2
FlowJo

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