Integrity of induced pluripotent stem cell (iPSC) derived megakaryocytes as assessed by genetic and transcriptomic analysis

Research output: Research - peer-reviewArticle

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.

LanguageEnglish (US)
Article numbere0167794
JournalPLoS ONE
Volume12
Issue number1
DOIs
StatePublished - Jan 1 2017

Fingerprint

megakaryocytes
transcriptomics
induced pluripotent stem cells
Stem cells
Induced Pluripotent Stem Cells
Megakaryocytes
Platelets
DNA
genotype
cells
Blood Platelets
Genotype
Structural integrity
Coagulation
Cell culture
Assays
Blood
Agglomeration
Genes
Chemical activation

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

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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.",
author = "Kai Kammers and Taub, {Margaret A.} and Ingo Ruczinski and Joshua Martin and Yanek, {Lisa R.} and Alyssa Frazee and Yongxing Gao and Dixie Hoyle and Nauder Faraday and Becker, {Diane M.} and Linzhao Cheng and Wang, {Zack Z.} and Leek, {Jeff T.} and Becker, {Lewis C.} and Mathias, {Rasika A.}",
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T1 - Integrity of induced pluripotent stem cell (iPSC) derived megakaryocytes as assessed by genetic and transcriptomic analysis

AU - Kammers,Kai

AU - Taub,Margaret A.

AU - Ruczinski,Ingo

AU - Martin,Joshua

AU - Yanek,Lisa R.

AU - Frazee,Alyssa

AU - Gao,Yongxing

AU - Hoyle,Dixie

AU - Faraday,Nauder

AU - Becker,Diane M.

AU - Cheng,Linzhao

AU - Wang,Zack Z.

AU - Leek,Jeff T.

AU - Becker,Lewis C.

AU - Mathias,Rasika A.

PY - 2017/1/1

Y1 - 2017/1/1

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