Integrity of Induced Pluripotent Stem Cell iPSC Derived Megakaryocytes as Assessed by Genetic and Transcriptomic AnalysisReportar como inadecuado




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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.



Autor: Kai Kammers, Margaret A. Taub, Ingo Ruczinski, Joshua Martin, Lisa R. Yanek, Alyssa Frazee, Yongxing Gao, Dixie Hoyle, Nauder Far

Fuente: http://plos.srce.hr/



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