Characterisation of iPSC derived Sensory neurons with Single Cell RNA-seq

Despite the rapid development of protocols to differentiate human pluripotent stem cells (hPSCs) into neurons, those used to produce sensory neurons remain difficult to replicate and result in heterogenous populations. Moreover, there is a growing clinical burden of chronic pain conditions, highlighting a need for relevant human cellular models. This study presents a hybrid differentiation method to produce nociceptive sensory neurons from hPSCs. Cell lines harbouring an inducible NEUROG2 construct were patterned towards precursors with small molecules, followed by NEUROG2 overexpression. We profilled cells with single cell RNA sequencing, which revealed populations of nociceptors, demonstrated by the expression of key developmental and fucntional genes including volatge gated sodium and TRP channels. Overall, this study represents an optimisation of existing methods to efficiently produce nociceptive sensory neurons and provides a tool for chronic pain research and the development of treatments. Overall design: Pools of neurons were processed and analysed at two time points in the differentiation protocol = D11 (precursors) and D40 (final time point). A population of iPSC derived cortical neurons produced with an iNeuron protocol was also analysed.