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. 2016 Sep 14;6(2):369–381. doi: 10.5966/sctm.2015-0424

Figure 6.

Figure 6

Contact‐mediated signaling between human iPSC‐derived sensory neurons and rat Schwann cells in coculture. (A): Images of myelin‐like segments along neuritic networks of human iPSC‐derived sensory neurons in coculture with rat Schwann cells for 14 days in medium supplemented with ascorbic acid. Double immunofluorescence for P0/neurofilament (Aa), MBP/neurofilament (Ab), and GALC/neurofilament (Ac). Cell nuclei were visualized with DAPI. Scale bars = 50 μm. (Ba): Western blot analysis for NRG1 type III in cell lysates prepared from iPSC‐derived iSNs alone or iSN‐Schwann cell cocultures for the indicated days under nonmyelinating conditions. Schwann cells were given this period to populate the neurites before induction of myelination with ascorbic acid supplementation. (Bb): Histogram showing NRG1 type III band density as normalized against the corresponding actin band density. NRG1 type III was detectable in extracts of iSNs and at increased levels in extracts of cocultures, leveling off by day 3 of coculture (Ba). ∗, p < .05, cocultures at days 1, 3, and 5 vs. iSNs alone. Abbreviations: DAPI, 4′,6‐diamidino‐2‐phenylindole; GALC, galactocerebroside; iPSC, induced pluripotent stem cell; iSN, induced sensory neuron; MBP, myelin basic protein; NRG1, neuregulin 1.