Rapid and efficient generation of transgene-free iPSC from a small volume of cryopreserved blood

Hongyan Zhou*, Hector Martinez, Bruce Sun, Aiqun Li, Matthew Zimmer, Elias Nicholas Katsanis, Erica Ellen Davis, Joanne Kurtzberg, Scott Lipnick, Scott Noggle, Mahendra Rao, Stephen Chang

*Corresponding author for this work

Research output: Contribution to journalArticle

16 Scopus citations

Abstract

Human peripheral blood and umbilical cord blood represent attractive sources of cells for reprogramming to induced pluripotent stem cells (iPSCs). However, to date, most of the blood-derived iPSCs were generated using either integrating methods or starting from T-lymphocytes that have genomic rearrangements thus bearing uncertain consequences when using iPSC-derived lineages for disease modeling and cell therapies. Recently, both peripheral blood and cord blood cells have been reprogrammed into transgene-free iPSC using the Sendai viral vector. Here we demonstrate that peripheral blood can be utilized formedium-throughput iPSC production without the need to maintain cell culture prior to reprogramming induction. Cell reprogramming can also be accomplished with as little as 3000 previously cryopreserved cord blood cells under feeder-free and chemically defined Xeno-free conditions that are compliant with standard Good Manufacturing Practice (GMP) regulations. The first iPSC colonies appear 2–3 weeks faster in comparison to previous reports. Notably, these peripheral blood- and cord bloodderived iPSCs are free of detectable immunoglobulin heavy chain (IGH) and T cell receptor (TCR) gene rearrangements, suggesting they did not originate from B- or T- lymphoid cells. The iPSCs are pluripotent as evaluated by the scorecard assay and in vitro multi lineage functional cell differentiation. Our data show that small volumes of cryopreserved peripheral blood or cord blood cells can be reprogrammed efficiently at a convenient, cost effective and scalable way. In summary, our method expands the reprogramming potential of limited or archived samples either stored at blood banks or obtained from pediatric populations that cannot easily provide large quantities of peripheral blood or a skin biopsy.

Original languageEnglish (US)
Article numberA004
Pages (from-to)652-665
Number of pages14
JournalStem Cell Reviews and Reports
Volume11
Issue number4
DOIs
StatePublished - Aug 1 2015

Keywords

  • Cell therapy
  • Cord blood
  • GMP
  • Genomic rearrangement
  • IPSC
  • Peripheral blood
  • Reprogramming
  • Sendai viral vector

ASJC Scopus subject areas

  • Cell Biology
  • Cancer Research

Fingerprint Dive into the research topics of 'Rapid and efficient generation of transgene-free iPSC from a small volume of cryopreserved blood'. Together they form a unique fingerprint.

  • Cite this

    Zhou, H., Martinez, H., Sun, B., Li, A., Zimmer, M., Katsanis, E. N., Davis, E. E., Kurtzberg, J., Lipnick, S., Noggle, S., Rao, M., & Chang, S. (2015). Rapid and efficient generation of transgene-free iPSC from a small volume of cryopreserved blood. Stem Cell Reviews and Reports, 11(4), 652-665. [A004]. https://doi.org/10.1007/s12015-015-9586-8