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. Author manuscript; available in PMC: 2013 Jan 1.
Published in final edited form as: Nat Biotechnol. 2012 Jul 1;30(7):715–720. doi: 10.1038/nbt.2249

Figure 2. LSB3i treated hPSCs accelerate via a neural crest intermediate into mature bipolar nociceptors with an action potential.

Figure 2

To monitor the emergence of neural crest stem cells, a transgenic SOX10∷GFP BAC hESC cell line was treated with (a) LSB, (b) LSB and CHIR99021, or (c) LSB3i. (d) SOX10∷GFP+ expression was accelerated and maximal expression (80% GFP+ by day 12) occurred earlier compared to LSB and CHIR99021 or LSB treatment alone. Sorted SOX10∷GFP+ cells gave rise to (e) ISL1 and (f) BRN3A positive neurons. (g) LSB3i neurons stain for glutamate. (h) Between days 8 and 14 RUNX1 expression is extinguished in some of the cells (higher expression in filled arrowhead, lower in empty arrowhead) and RET upregulates. (i) By day 15, mature neurons express peripherin, and after 1 month, (j) neuronal cell bodies arrange as clusters positive for (k) Substance P and (l) CGRP. Scale bars for (a-g, i-m) are 100 μm and 50 μm for (h).