TABLE 4.

Troubleshooting table.

Step	Problem	Possible reason	Solution
18	EBs do not attach well	EBs are too big or too small; they contain too many dead cells. Reduce the time for treatment with dispase	Avoid overtrituration of EBs. Gently pipette the EBs during medium changes to remove dead cells from the surface
	Low neural induction efficiency of hiPSCs	Variation in neural induction of iPSCs	Treat EBs with SB 431542 (10 μM) and DMH1 (10 μM) for the first 6 d
21	The medium becomes orange or yellow too quickly	Too many cells in one flask	Cells should be fed every day or split into two flasks. An acidic environment may increase cell death
26	Large spheres	Cell proliferation	Triturate the spheres with a glass Pasteur pipette (Fig. 6)