Figure 1. Schematic for the STEMdiff™ Sensory Neuron Culture System Protocol

Sensory neuron precursors can be generated in 6 days from hPSC-derived neural crest cells. For the generation of neural crest cells, see documentation for STEMdiff™ Neural Crest Differentiation Kit (Catalog #08610). For the maturation of sensory neuron precursors to sensory neurons, see the PIS.

Figure 2. STEMdiff™ Sensory Neuron Kits Promote Differentiation Across Multiple Embryonic Stem and Induced Pluripotent Stem Cell Lines

NCCs generated from hPSCs in mTeSR™ Plus using the STEMdiff™ Neural Crest Differentiation Kit were differentiated and matured to sensory neurons using the STEMdiff™ Sensory Neuron Differentiation and Maturation Kits. (A) Sensory neurons were generated after hPSC-derived NCCs were cultured with the STEMdiff™ Sensory Neuron Differentiation Kit for 6 days and then the STEMdiff™ Sensory Neuron Maturation Kit for 6 days. The resulting cultures contain a population of cells expressing sensory neuron markers peripherin (green) and BRN3A (red) along with (B) neuronal marker class III β-tubulin (TUJ1, red). (C) Midbrain neuron controls generated with STEMdiff™ Midbrain Neuron Differentiation and Maturation Kits do not have detectable peripherin (green) or BRN3A (red) expression, although they express (D) neuronal marker class III β-tubulin (TUJ1, red). Nuclei are labeled with DAPI (blue). Human ES and iPS cell lines were maintained in either mTeSR™1, TeSR™-E8™, or mTeSR™ Plus and differentiated with STEMdiff™ Neural Crest Differentiation Kit, followed by STEMdiff™ Sensory Neuron Differentiation and Maturation Kits. The percentage expression of (E) BRN3A+ and (F) TUJ1+ cells in the resulting cultures was quantified. This differentiation generated BRN3A+ sensory neurons (25.3% ± 6.9%, mean ± SEM; n=7 cell lines, 3 - 23 replicates per condition) that expressed neuronal marker class III β-tubulin (TUJ1; 90.3% ± 4.1%, mean ± SEM; n=4 cell lines, 3 - 12 replicates per condition). Numbers are % positive over total DAPI in a tiled image. NCCs = neural crest cells; hPSCs = human pluripotent stem cells; ES = embryonic stem; iPS = induced pluripotent stem