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. Author manuscript; available in PMC: 2018 Nov 19.
Published in final edited form as: Nat Protoc. 2018 Feb 22;13(3):565–580. doi: 10.1038/nprot.2017.152

Figure 2 |.

Figure 2 |

Schematic diagram of brain region–specific organoid protocols. The protocols start with enzymatically detaching hiPSC colonies cultured on MEF at Day 0 to generate EBs (Steps 1–7 of the PROCEDURE). From here, the protocols diverge for the procedures to differentiate forebrain (Steps 9–32of the PROCEDURE), midbrain (Box 2) and hypothalamus (Box 3) organoids, respectively. The forebrain organoid protocol involves two unique procedures for embedding EBs in Matrigel at Day 7 and dissociating from Matrigel at Day 14, detailed in Steps 11–25. Finally, the procedures converge with culturing of organoids in the SpinΩ bioreactor, as described in Steps 26–30.