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Cellular and Molecular Life Sciences

, Volume 72, Issue 23, pp 4671–4680 | Cite as

Selective cell targeting and lineage tracing of human induced pluripotent stem cells using recombinant avian retroviruses

  • Laura Hildebrand
  • Petra Seemann
  • Andreas Kurtz
  • Jochen Hecht
  • Jörg Contzen
  • Manfred Gossen
  • Harald StachelscheidEmail author
Research Article

Abstract

Human induced pluripotent stem cells (hiPSC) differentiate into multiple cell types. Selective cell targeting is often needed for analyzing gene function by overexpressing proteins in a distinct population of hiPSC-derived cell types and for monitoring cell fate in response to stimuli. However, to date, this has not been possible, as commonly used viruses enter the hiPSC via ubiquitously expressed receptors. Here, we report for the first time the application of a heterologous avian receptor, the tumor virus receptor A (TVA), to selectively transduce TVA+ cells in a mixed cell population. Expression of the TVA surface receptor via genetic engineering renders cells susceptible for infection by avian leucosis virus (ALV). We generated hiPSC lines with this stably integrated, ectopic TVA receptor gene that expressed the receptor while retaining pluripotency. The undifferentiated hiPSCTVA+ as well as their differentiating progeny could be infected by recombinant ALV (so-called RCAS virus) with high efficiency. Due to incomplete receptor blocking, even sequential infection of differentiating or undifferentiated TVA+ cells was possible. In conclusion, the TVA/RCAS system provides an efficient and gentle gene transfer system for hiPSC and extends our possibilities for selective cell targeting and lineage tracing studies.

Keywords

Induced pluripotent stem cells TVA ALV Selective cell targeting Lineage tracing 

Notes

Acknowledgments

We thank Mareen Schmidt-von Kegler and Maria Walther as well as the Berlin Institute of Health (BIH) Stem Cell Core Facility for excellent technical assistance and the people from the laboratory of Prof. Petra Seemann and Dr. Andreas Kurtz for helpful discussions regarding the study. Additionally, we thank Dr. Stephen Hughes for kindly providing us with the RCAS and RCAN vectors and Dr. Ansgar Petersen for help at the two-photon microscope. We appreciate the financial support and provision of laboratory facilities by Prof. Petra Reinke. Laura Hildebrand is a member of the DFG funded Berlin-Brandenburg School for Regenerative Therapies GSC 203. This work was supported by the grant 1315848A for the Berlin-Brandenburg Center for Regenerative Therapies (BCRT) by the Initiative and Networking Fund of the Helmholtz Association, Germany.

Conflict of interest

All authors state that they have no conflicts of interest.

Supplementary material

18_2015_1957_MOESM1_ESM.jpg (158 kb)
Supplementary material 1 (JPEG 157 kb) Supplemental Figure1: Confirmation of trilineage differentiation potential of Clone 1 hiPSCTVA+ Undirected differentiation of the hiPSCTVA+ as embryoid bodies (EBs) in two different media showed upregulation of markers for the three germ layers, while markers for pluripotency were downregulated
18_2015_1957_MOESM2_ESM.docx (17 kb)
Supplementary material 2 (DOCX 17 kb)
18_2015_1957_MOESM3_ESM.docx (13 kb)
Supplementary material 3 (DOCX 12 kb)

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Copyright information

© Springer Basel 2015

Authors and Affiliations

  • Laura Hildebrand
    • 1
    • 2
  • Petra Seemann
    • 1
    • 2
  • Andreas Kurtz
    • 1
    • 2
    • 4
  • Jochen Hecht
    • 1
    • 2
    • 6
  • Jörg Contzen
    • 2
    • 3
  • Manfred Gossen
    • 2
    • 3
  • Harald Stachelscheid
    • 1
    • 2
    • 5
    Email author
  1. 1.Charité-Universitätsmedizin BerlinBerlinGermany
  2. 2.Berlin-Brandenburg Center for Regenerative Therapies (BCRT)BerlinGermany
  3. 3.Helmholtz-Zentrum Geesthacht (HZG), Institute of Biomaterial ScienceTeltowGermany
  4. 4.College of Veterinary Medicine and Research Institute for Veterinary ScienceSeoul National UniversitySeoulRepublic of Korea
  5. 5.Berlin Institute of Health-Stem Cell Core FacilityBerlinGermany
  6. 6.Max Planck Institute for Molecular GeneticsBerlinGermany

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