Protocol for Differentiation of Human iPSCs into Pulmonary Neuroendocrine Cells

Summary

Pulmonary neuroendocrine cells (PNECs) are sensory cells within the lung airway epithelia. Here, we provide a detailed protocol for generating induced PNECs (iPNECs) from human induced pluripotent stem cells (iPSCs). The cellular and molecular profile of iPNECs resembles primary human PNECs. Primary human PNECs are exceedingly rare, comprising only 1% of the adult lung. Therefore, a self-renewing source of patient-specific iPNECs facilitates the creation of reproducible human cellular models to study lung diseases characterized by PNEC dysfunction.

For complete details on the use and execution of this protocol, please refer to Hor et al. (2020).

Graphical Abstract

Highlights

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  Efficient generation of iPNECs from human iPSCs

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  Air-liquid-interface culture augments iPNEC specification

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  Continuous Notch inhibition increases iPNEC specification

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  iPNEC gene expression signature is highly concordant with primary human fetal PNECs

Abstract

Pulmonary neuroendocrine cells (PNECs) are sensory cells within the lung airway epithelia. Here, we provide a detailed protocol for generating induced PNECs (iPNECs) from human induced pluripotent stem cells (iPSCs). The cellular and molecular profile of iPNECs resembles primary human PNECs. Primary human PNECs are exceedingly rare, comprising only 1% of the adult lung. Therefore, a self-renewing source of patient-specific iPNECs facilitates the creation of reproducible human cellular models to study lung diseases characterized by PNEC dysfunction.