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

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.

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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.

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All authors state that they have no conflicts of interest.

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Correspondence to Harald Stachelscheid.

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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

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Supplementary material 3 (DOCX 12 kb)

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Hildebrand, L., Seemann, P., Kurtz, A. et al. Selective cell targeting and lineage tracing of human induced pluripotent stem cells using recombinant avian retroviruses. Cell. Mol. Life Sci. 72, 4671–4680 (2015). https://doi.org/10.1007/s00018-015-1957-4

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Keywords

  • Induced pluripotent stem cells
  • TVA
  • ALV
  • Selective cell targeting
  • Lineage tracing