Abstract
The generation of human sensory neurons by directed differentiation of pluripotent stem cells opens new opportunities for investigating the biology of pain. The inability to generate this cell type has meant that up until now their study has been reliant on the use of rodent models. Here, we use a combination of population and single-cell techniques to perform a detailed molecular, electrophysiological, and pharmacological phenotyping of sensory neurons derived from human embryonic stem cells. We describe the evolution of cell populations over 6 weeks of directed differentiation; a process that results in the generation of a largely homogeneous population of neurons that are both molecularly and functionally comparable to human sensory neurons derived from mature dorsal root ganglia. This work opens the prospect of using pluripotent stem-cell–derived sensory neurons to study human neuronal physiology and as in vitro models for drug discovery in pain and sensory disorders.
Highlights
The in vitro generation of differentiated cells from pluripotent stem cells is a key goal of regenerative medicine
Generation of human pluripotent stem cells (hPSC)-sensory from small molecule inhibition of BMP, GSK3, γ-secretase, vascular endothelial growth factor receptor, and fibroblast growth factor receptor followed by growth in neurotrophin media We have previously reported generation of functional sensorylike neurons from human embryonic stem cell cells using a Population Single cell
We found that our hPSC-sensory protocol was capable of effectively neuralizing pluripotent cells, driving them through a neuroectodermal fate followed by neural crest and the formation of a distinct nociceptor phenotype; a process that broadly recapitulates the in vivo developmental process.[18]
Summary
The in vitro generation of differentiated cells from pluripotent stem cells is a key goal of regenerative medicine. Such cells can be used in place of human tissue or animal models for disease modeling, drug screening, and even cell replacement therapies.[1] A major challenge, addressed here, is improving the level of molecular and cellular understanding of the in vitro differentiation process and in depth functional characterization of the terminally differentiated cells produced. A relative scarcity of protocols exist that describe the derivation of sensory neurons of the peripheral nervous system.[9,10] The lack of access to this tissue has limited our understanding of its development and the physiology of pain in humans
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