Generation of transgene-free iPSC lines from human normal and neoplastic blood cells using episomal vectors

Kejin Hu, Igor Slukvin

Research output: Chapter in Book/Report/Conference proceedingConference contributionpeer-review

21 Scopus citations


Human induced pluripotent stem cells (iPSCs) have become an important tool for modeling human diseases and are considered a potential source of therapeutic cells. Original methods for iPSC generation use fibroblasts as a cell source for reprogramming and retroviral vectors as a delivery method of the reprogramming factors. However, fibroblasts require extended time for expansion and viral delivery of transgenes results in the integration of vector sequences into the genome which is a source of potential insertion mutagenesis, residual expressions, and reactivation of transgenes during differentiation. Here, we provide a detailed protocol for the efficient generation of transgene-free iPSC lines from human bone marrow and cord blood cells with a single transfection of non-integrating episomal plasmids. This method uses mononuclear bone marrow and cord blood cells, and makes it possible to generate transgene-free iPSCs 1-3 weeks faster than previous methods of reprogramming with fibroblasts. Additionally, we show that this approach can be used for efficient reprogramming of chronic myeloid leukemia cells.

Original languageEnglish
Title of host publicationPluripotent Stem Cells
Subtitle of host publicationMethods and Protocols
EditorsUma Lakshmipathy, Mohan Vemuri
Number of pages14
StatePublished - 2013

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745


  • Chronic myeloid leukemia
  • Cord blood
  • Episomal plasmids
  • Epstein-Barr virus
  • Human bone marrow
  • Induced pluripotent stem cells
  • Reprogramming


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