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dc.contributor.authorGaritaonandia, I
dc.contributor.authorAmir, H
dc.contributor.authorBoscolo, FS
dc.contributor.authorWambua, GK
dc.contributor.authorSchultheisz, HL
dc.contributor.authorSabatini, K
dc.contributor.authorMorey, R
dc.contributor.authorWaltz, S
dc.contributor.authorWang, Y-C
dc.contributor.authorTran, H
dc.contributor.authorLeonardo, TR
dc.contributor.authorNazor, K
dc.contributor.authorSlavin, I
dc.contributor.authorLynch, C
dc.contributor.authorLi, Y
dc.contributor.authorColeman, R
dc.contributor.authorRomero, IG
dc.contributor.authorAltun, G
dc.contributor.authorReynolds, D
dc.contributor.authorDalton, S
dc.contributor.authorParast, M
dc.contributor.authorLoring, JF
dc.contributor.authorLaurent, LC
dc.date.accessioned2020-12-18T04:28:48Z
dc.date.available2020-12-18T04:28:48Z
dc.date.issued2015-02-25
dc.identifierpii: PONE-D-14-46378
dc.identifier.citationGaritaonandia, I., Amir, H., Boscolo, F. S., Wambua, G. K., Schultheisz, H. L., Sabatini, K., Morey, R., Waltz, S., Wang, Y. -C., Tran, H., Leonardo, T. R., Nazor, K., Slavin, I., Lynch, C., Li, Y., Coleman, R., Romero, I. G., Altun, G., Reynolds, D. ,... Laurent, L. C. (2015). Increased Risk of Genetic and Epigenetic Instability in Human Embryonic Stem Cells Associated with Specific Culture Conditions. PLOS ONE, 10 (2), https://doi.org/10.1371/journal.pone.0118307.
dc.identifier.issn1932-6203
dc.identifier.urihttp://hdl.handle.net/11343/256211
dc.description.abstractThe self-renewal and differentiation capacities of human pluripotent stem cells (hPSCs) make them a promising source of material for cell transplantation therapy, drug development, and studies of cellular differentiation and development. However, the large numbers of cells necessary for many of these applications require extensive expansion of hPSC cultures, a process that has been associated with genetic and epigenetic alterations. We have performed a combinatorial study on both hESCs and hiPSCs to compare the effects of enzymatic vs. mechanical passaging, and feeder-free vs. mouse embryonic fibroblast feeder substrate, on the genetic and epigenetic stability and the phenotypic characteristics of hPSCs. In extensive experiments involving over 100 continuous passages, we observed that both enzymatic passaging and feeder-free culture were associated with genetic instability, higher rates of cell proliferation, and persistence of OCT4/POU5F1-positive cells in teratomas, with enzymatic passaging having the stronger effect. In all combinations of culture conditions except for mechanical passaging on feeder layers, we noted recurrent deletions in the genomic region containing the tumor suppressor gene TP53, which was associated with decreased mRNA expression of TP53, as well as alterations in the expression of several downstream genes consistent with a decrease in the activity of the TP53 pathway. Among the hESC cultures, we also observed culture-associated variations in global gene expression and DNA methylation. The effects of enzymatic passaging and feeder-free conditions were also observed in hiPSC cultures. Our results highlight the need for careful assessment of the effects of culture conditions on cells intended for clinical therapies.
dc.languageEnglish
dc.publisherPUBLIC LIBRARY SCIENCE
dc.titleIncreased Risk of Genetic and Epigenetic Instability in Human Embryonic Stem Cells Associated with Specific Culture Conditions
dc.typeJournal Article
dc.identifier.doi10.1371/journal.pone.0118307
melbourne.affiliation.departmentSchool of BioSciences
melbourne.source.titlePLoS One
melbourne.source.volume10
melbourne.source.issue2
dc.rights.licenseCC BY
melbourne.elementsid1215123
melbourne.contributor.authorGallego Romero, Irene
dc.identifier.eissn1932-6203
melbourne.accessrightsOpen Access


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