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Towards maturation of human otic hair cell–like cells in pluripotent stem cell–derived organoid transplants

Abstract

Human otic organoids generated from pluripotent stem cells (PSCs) provide a promising platform for modeling, drug testing, and cell-based therapies of inner ear diseases. However, providing the appropriate niche that resembles inner ear development and its vasculature to generate otic organoids is less conspicuous. Here, we devised a strategy to enhance maturation of otic progenitor cells toward human hair cell–like cells (HCLCs) by assembling three-dimensional (3D) otic organoids that contain human PSC-derived otic cells, endothelial cells, and mesenchymal stem cells (MSCs). Heterotopic implantation of otic organoids, designated as grafted otic organoids (GOs), in ex ovo chick embryo chorioallantoic membrane (CAM) stimulated maturation of the HCLCs. Functional analysis revealed the presence of voltage-gated potassium currents without detectable sodium currents in these cells in the GOs. Our results demonstrated that implantation of 3D heterotypic cell mixtures of otic organoids improved maturation of human HCLCs. This GO-derived HCLCs could be an attractive source for drug discovery and other biomedical applications.

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taken from GO slice indicates recording from a HCLC. Inset showed the morphology of the cell that was chosen for electrophysiological recording. Scale bar 20 μm. Inset 10 µm (d). A sample of voltage clamp signals recorded from GO cells using the same protocol as b. The cells exhibited a detectable outward current when the membrane was depolarized from − 90 mV to potentials above + 40 mV. e Quantitative current–voltage relation of maximal outward current elicited by applying the depolarizing voltage pulses from −60 to + 30 mV. GO, grafted otic organoid; VO, in vitro otic organoid

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Acknowledgements

We would like to express our appreciation to Professor Allen F. Ryan for his critical input and to Paria Pooyan for helping us to improve our schematic images.

Funding

This study was funded by a grant from Royan Institute and the Cognitive Science and Technologies Council of Iran.

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Authors

Contributions

F.M. designed the study, performed most of the experiments, discussed the results, and wrote the manuscript. A.S. performed and analyzed the electrophysiology results. N.H. contributed to the design and analysis of the experiments. S.Y. and S.N.H. contributed to the heterotopic implantation of the otic organoids in the ex ovo chick embryo chorioallantoic membrane (CAM). S.N. contributed to the design of the study and provided financial support. H.B. provided financial and administrative support, designed and analyzed the experiments, interpreted and discussed the results, and approved the manuscript. All the authors reviewed and confirmed the manuscript before submission.

Corresponding author

Correspondence to Hossein Baharvand.

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This study and all animal experiments were approved by the Institutional Review Board and Ethical Committee of Royan Institute (Tehran, Iran).

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The authors declare no competing interests.

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Moeinvaziri, F., Shojaei, A., Haghparast, N. et al. Towards maturation of human otic hair cell–like cells in pluripotent stem cell–derived organoid transplants. Cell Tissue Res (2021). https://doi.org/10.1007/s00441-021-03510-y

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Keywords

  • CAM transplantation
  • Organoid
  • Human otic differentiation
  • Hair cell-like cell
  • Maturation