Table 2.

Troubleshooting table.

Step	Problem	Possible reason	Solution
1	Low cell survival post-thaw.	Cell death during thaw.	Do not let the vial of cells completely thaw in the water bath.
2	#x02013;5	hPSCs are not reaching confluency at the predicted time.	hPSCs are too big or too small; they contain too many dead cells.	The passage ratio can be changed to increase cell density. Gently pipette the iPSCs during passage and medium changes.
18	The medium becomes 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.
20	Large spheres.	Cell proliferation.	Extend the processing time of Accutase Solution.
21	#x02013;23	Cell layers are detaching from plate during differentiation.	Inefficient Matrigel coating.	Use fresh Matrigel and make sure to coat for 24 h. Take out the Matrigel-coated tissue culture plates from the freezer. Leave the plate at 37°C in an incubator for 1 h.
24	#x02013;31	Cell layer peeling during staining.	Cell layers are releasing during fixation and wash steps.	Fix for 1 h instead of 30 min. During wash steps, a transfer pipette may be used to carefully exchange liquids.