title: Integrity of Induced Pluripotent Stem Cell (iPSC) Derived Megakaryocytes as Assessed by Genetic and Transcriptomic Analysis.
abstract: Previously, we have described our feeder-free, xeno-free approach to generate megakaryocytes (MKs) in culture from human induced pluripotent stem cells (iPSCs). Here, we focus specifically on the integrity of these MKs using: (1) genotype discordance between parent cell DNA to iPSC cell DNA and onward to the differentiated MK DNA; (2) genomic structural integrity using copy number variation (CNV); and (3) transcriptomic signatures of the derived MK lines compared to the iPSC lines. We detected a very low rate of genotype discordance; estimates were 0.0001%-0.01%, well below the genotyping error rate for our assay (0.37%). No CNVs were generated in the iPSCs that were subsequently passed on to the MKs. Finally, we observed highly biologically relevant gene sets as being upregulated in MKs relative to the iPSCs: platelet activation, blood coagulation, megakaryocyte development, platelet formation, platelet degranulation, and platelet aggregation. These data strongly support the integrity of the derived MK lines.
authors.affiliation: Division of Biostatistics and Bioinformatics, Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America.|Department of Biostatistics, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, United States of America.
authors.forename: Kai
authors.initials: K
authors.lastname: Kammers
authors.affiliation: Department of Biostatistics, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, United States of America.
authors.forename: Margaret A
authors.initials: MA
authors.lastname: Taub
authors.affiliation: Department of Biostatistics, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, United States of America.
authors.forename: Ingo
authors.initials: I
authors.lastname: Ruczinski
authors.affiliation: The GeneSTAR Research Program, Johns Hopkins School of Medicine, Baltimore, Maryland, United States of America.
authors.forename: Joshua
authors.initials: J
authors.lastname: Martin
authors.affiliation: The GeneSTAR Research Program, Johns Hopkins School of Medicine, Baltimore, Maryland, United States of America.
authors.forename: Lisa R
authors.initials: LR
authors.lastname: Yanek
authors.affiliation: Department of Biostatistics, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, United States of America.
authors.forename: Alyssa
authors.initials: A
authors.lastname: Frazee
authors.affiliation: Division of Hematology and Institute for Cell Engineering, Johns Hopkins School of Medicine, Baltimore, Maryland, United States of America.
authors.forename: Yongxing
authors.initials: Y
authors.lastname: Gao
authors.affiliation: Division of Hematology and Institute for Cell Engineering, Johns Hopkins School of Medicine, Baltimore, Maryland, United States of America.
authors.forename: Dixie
authors.initials: D
authors.lastname: Hoyle
authors.affiliation: The GeneSTAR Research Program, Johns Hopkins School of Medicine, Baltimore, Maryland, United States of America.
authors.forename: Nauder
authors.initials: N
authors.lastname: Faraday
authors.affiliation: The GeneSTAR Research Program, Johns Hopkins School of Medicine, Baltimore, Maryland, United States of America.
authors.forename: Diane M
authors.initials: DM
authors.lastname: Becker
authors.affiliation: Division of Hematology and Institute for Cell Engineering, Johns Hopkins School of Medicine, Baltimore, Maryland, United States of America.
authors.forename: Linzhao
authors.initials: L
authors.lastname: Cheng
authors.affiliation: Division of Hematology and Institute for Cell Engineering, Johns Hopkins School of Medicine, Baltimore, Maryland, United States of America.
authors.forename: Zack Z
authors.initials: ZZ
authors.lastname: Wang
authors.affiliation: Department of Biostatistics, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, United States of America.
authors.forename: Jeff T
authors.initials: JT
authors.lastname: Leek
authors.affiliation: The GeneSTAR Research Program, Johns Hopkins School of Medicine, Baltimore, Maryland, United States of America.
authors.forename: Lewis C
authors.initials: LC
authors.lastname: Becker
authors.affiliation: The GeneSTAR Research Program, Johns Hopkins School of Medicine, Baltimore, Maryland, United States of America.
authors.forename: Rasika A
authors.initials: RA
authors.lastname: Mathias
mesh_terms: D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D008533:Megakaryocytes / Q000166:cytology* / Q000378:metabolism; D059467:Transcriptome