Open AccessIntegrity of Induced Pluripotent Stem Cell (iPSC) Derived Megakaryocytes as Assessed by Genetic and Transcriptomic Analysis
Author(s) -
Kai Kammers,
Margaret A. Taub,
Ingo Ruczinski,
Joshua Martin,
Lisa R. Yanek,
Alyssa C. Frazee,
Yongxing Gao,
Dixie L. Hoyle,
Nauder Faraday,
Diane M. Becker,
Linzhao Cheng,
Zack Z. Wang,
Jeff Leek,
Lewis C. Becker,
Rasika A. Mathias
Publication year - 2017
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0167794
Subject(s) - induced pluripotent stem cell , megakaryocyte , biology , transcriptome , stem cell , microbiology and biotechnology , platelet , genotyping , genotype , gene , genetics , haematopoiesis , embryonic stem cell , gene expression , immunology
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.