Protocol for inducing dorsal spinal sensory interneurons from mouse embryonic stem cell-derived neuromesodermal progenitors

Here, we present a protocol comprising two differentiation techniques to derive dorsal sensory spinal interneurons (dIs) from mouse embryonic stem cells (mESCs) through a neuromesodermal progenitor (NMP) precursor. First, we describe the steps for using retinoic acid (RA) to induce dI4s, dI5s, and dI6s, which mediate pain, itch, touch, and sensorimotor integration. Second, we detail the procedures for generating dI1s, dI2s, and dI3s, which mediate proprioception and mechanosensation, using RA together with bone morphogenetic protein (BMP) 4. For complete details on the use and execution of this protocol, please refer to Gupta et al.¹.