Blood cell-derived induced pluripotent stem cells free of reprogramming factors generated by Sendai viral vectors

Stem Cells Transl Med. 2013 Aug;2(8):558-66. doi: 10.5966/sctm.2013-0006. Epub 2013 Jul 11.

Abstract

The discovery of induced pluripotent stem cells (iPSCs) holds great promise for regenerative medicine since it is possible to produce patient-specific pluripotent stem cells from affected individuals for potential autologous treatment. Using nonintegrating cytoplasmic Sendai viral vectors, we generated iPSCs efficiently from adult mobilized CD34⁺ and peripheral blood mononuclear cells. After 5-8 passages, the Sendai viral genome could not be detected by real-time quantitative reverse transcription-polymerase chain reaction. Using the spin embryoid body method, we showed that these blood cell-derived iPSCs could efficiently be differentiated into hematopoietic stem and progenitor cells without the need of coculture with either mouse or human stromal cells. We obtained up to 40% CD34⁺ of which ~25% were CD34⁺/CD43⁺ hematopoietic precursors that could readily be differentiated into mature blood cells. Our study demonstrated a reproducible protocol for reprogramming blood cells into transgene-free iPSCs by the Sendai viral vector method. Maintenance of the genomic integrity of iPSCs without integration of exogenous DNA should allow the development of therapeutic-grade stem cells for regenerative medicine.

Keywords: CD34+; Differentiation; Hematopoietic progenitors; Reprogramming.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adult
  • Animals
  • Antigens, CD34 / metabolism
  • Blood Cells / cytology*
  • Cell Differentiation / genetics
  • Cellular Reprogramming* / genetics
  • Female
  • Gene Expression Profiling
  • Genetic Vectors / metabolism*
  • Hematopoiesis / genetics
  • Hematopoietic Stem Cells / cytology
  • Hematopoietic Stem Cells / metabolism
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism
  • Male
  • Mice
  • Mice, Inbred NOD
  • Sendai virus / genetics*
  • Transcription Factors / metabolism*
  • Transgenes / genetics

Substances

  • Antigens, CD34
  • Transcription Factors