Messenger RNA- Versus Retrovirus-Based Induced Pluripotent Stem Cell Reprogramming Strategies: Analysis of Genomic Integrity

Autor: Noushin Dianat, Inmaculada Moreno-Gimeno, Eléanor Luce, Anne Dubart-Kupperschmitt, Gérard Tachdjian, Joëlle Marie, Clara Steichen, Lucie Tosca, Jérome Maluenda, Deborah J. Burks, Christophe Desterke, Judith Melki, Sylvie Goulinet-Mainot, Sarah Awan-Toor, Anne Weber
Jazyk: angličtina
Rok vydání: 2014
Předmět:
DNA Copy Number Variations
Genotype
Cellular differentiation
Genetic Vectors
Induced Pluripotent Stem Cells
iPSCs
Computational biology
Transfection
Polymorphism
Single Nucleotide
Regenerative medicine
03 medical and health sciences
0302 clinical medicine
Retrovirus
Humans
RNA
Messenger
Copy-number variation
Induced pluripotent stem cell
Cells
Cultured
Oligonucleotide Array Sequence Analysis
Embryonic Stem Cells/Induced Pluripotent Stem (iPS) Cells
030304 developmental biology
Genetics
0303 health sciences
biology
Gene Expression Profiling
Gene Expression Regulation
Developmental
Cell Differentiation
Genome integrity
Hepatic differentiation
Cell Biology
General Medicine
Fibroblasts
Cellular Reprogramming
biology.organism_classification
3. Good health
Gene expression profiling
Phenotype
Retroviridae
030220 oncology & carcinogenesis
Hepatocytes
SNP/CNV analysis
Reprogramming
Transcription Factors
Developmental Biology
SNP array
mRNA reprogramming
Zdroj: Stem Cells Translational Medicine
r-CIPF: Repositorio Institucional Producción Científica del Centro de Investigación Principe Felipe (CIPF)
Centro de Investigación Principe Felipe (CIPF)
r-CIPF. Repositorio Institucional Producción Científica del Centro de Investigación Principe Felipe (CIPF)
instname
ISSN: 2157-6580
2157-6564
DOI: 10.5966/sctm.2013-0158
Popis: The use of synthetic messenger RNAs to generate human induced pluripotent stem cells (iPSCs) is particularly appealing for potential regenerative medicine applications, because it overcomes the common drawbacks of DNA-based or virus-based reprogramming strategies, including transgene integration in particular. We compared the genomic integrity of mRNA-derived iPSCs with that of retrovirus-derived iPSCs generated in strictly comparable conditions, by single-nucleotide polymorphism (SNP) and copy number variation (CNV) analyses. We showed that mRNA-derived iPSCs do not differ significantly from the parental fibroblasts in SNP analysis, whereas retrovirus-derived iPSCs do. We found that the number of CNVs seemed independent of the reprogramming method, instead appearing to be clone-dependent. Furthermore, differentiation studies indicated that mRNA-derived iPSCs differentiated efficiently into hepatoblasts and that these cells did not load additional CNVs during differentiation. The integration-free hepatoblasts that were generated constitute a new tool for the study of diseased hepatocytes derived from patients' iPSCs and their use in the context of stem cell-derived hepatocyte transplantation. Our findings also highlight the need to conduct careful studies on genome integrity for the selection of iPSC lines before using them for further applications.
Databáze: OpenAIRE
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