Generation of induced pluripotent stem cells (iPSCs) from a Bernard-Soulier syndrome patient carrying a W71R mutation in the GPIX gene

Stem Cell Res. 2016 May;16(3):692-5. doi: 10.1016/j.scr.2016.04.013. Epub 2016 Apr 15.

Abstract

We generated an induced pluripotent stem cell (iPSC) line from a Bernard-Soulier Syndrome (BSS) patient carrying the mutation p.Trp71Arg in the GPIX locus (BSS1-PBMC-iPS4F4). Peripheral blood mononuclear cells (PBMCs) were reprogrammed using heat sensitive non-integrative Sendai viruses containing the reprogramming factors Oct3/4, SOX2, KLF4 and c-MYC. Successful silencing of the exogenous reprogramming factors was checked by RT-PCR. Characterization of BSS1-PBMC-iPS4F4 included mutation analysis of GPIX locus, Short Tandem Repeats (STR) profiling, alkaline phosphatase enzymatic activity, analysis of conventional pluripotency-associated factors at mRNA and protein level and in vivo differentiation studies. BSS1-PBMC-iPS4F4 will provide a powerful tool to study BSS.

MeSH terms

  • Animals
  • Bernard-Soulier Syndrome / metabolism
  • Bernard-Soulier Syndrome / pathology*
  • Cell Differentiation
  • Cells, Cultured
  • Cellular Reprogramming
  • Female
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism
  • Induced Pluripotent Stem Cells / transplantation
  • Karyotype
  • Kruppel-Like Factor 4
  • Leukocytes, Mononuclear / cytology
  • Leukocytes, Mononuclear / metabolism
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • Mutation
  • Platelet Glycoprotein GPIb-IX Complex / genetics*
  • Teratoma / metabolism
  • Teratoma / pathology
  • Transcription Factors / genetics
  • Transcription Factors / metabolism

Substances

  • KLF4 protein, human
  • Klf4 protein, mouse
  • Kruppel-Like Factor 4
  • Platelet Glycoprotein GPIb-IX Complex
  • Transcription Factors
  • adhesion receptor