Increased risk of genetic and epigenetic instability in human embryonic stem cells associated with specific culture conditions

Autor: Kristopher L. Nazor, Irene Gallego Romero, Louise C. Laurent, Ronald Coleman, Ileana Slavin, Robert Morey, Gerald K. Wambua, Francesca S. Boscolo, Stephen Dalton, Heather L. Schultheisz, Yingchun Li, Shannon Waltz, Hadar Amir, David Reynolds, Trevor R. Leonardo, Karen Sabatini, Gulsah Altun, Ibon Garitaonandia, Mana M. Parast, Jeanne F. Loring, Ha Tran, Yu-Chieh Wang, Candace L. Lynch
Přispěvatelé: Roh, Tae-Young
Jazyk: angličtina
Rok vydání: 2015
Předmět:
Time Factors
Cellular differentiation
Human Embryonic Stem Cells
Chromosomes
Human
Pair 20
Cell Culture Techniques
Cell Transformation
Regenerative Medicine
Epigenesis
Genetic
0302 clinical medicine
Chromosome Duplication
Pair 12
Cell Self Renewal
Induced pluripotent stem cell
Cells
Cultured
0303 health sciences
Cultured
Genome
Multidisciplinary
Cell Differentiation
Single Nucleotide
3. Good health
Cell biology
Cell Transformation
Neoplastic
Phenotype
DNA methylation
Medicine
Chromosome Deletion
Research Article
Human
Biotechnology
Pluripotent Stem Cells
Tumor suppressor gene
General Science & Technology
Cells
Science
Biology
Polymorphism
Single Nucleotide
Chromosomes
Genomic Instability
Cell Line
03 medical and health sciences
Genetic
Genetics
Humans
Epigenetics
Polymorphism
Stem Cell Research - Embryonic - Human
030304 developmental biology
Chromosome Aberrations
Neoplastic
Chromosomes
Human
Pair 12
Genome
Human
Pair 17
Gene Expression Profiling
DNA Methylation
Stem Cell Research
Embryonic stem cell
Molecular biology
Gene expression profiling
Cell culture
Generic health relevance
Tumor Suppressor Protein p53
Pair 20
030217 neurology & neurosurgery
Chromosomes
Human
Pair 17
Epigenesis
Zdroj: PLoS ONE, Vol 10, Iss 2, p e0118307 (2015)
PloS one, vol 10, iss 2
PLoS ONE
ISSN: 1932-6203
Popis: The 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.
Databáze: OpenAIRE
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