Table 5.

Troubleshooting table

Step	Problem	Possible reason	Solution
6	Not enough dissociation of cells	Cell density is too high	Start experiment with a lower density of hPSCs.
	Not enough agitation of cells	Tap side of the plate.
17	Low cell density	Low concentration of plated cells	Check plating cell density.
		Low attachment of the plated cells	Check thiazovivin concentration and expiry date.
			Check protein concentration and expiry data of Geltrex.
		Poor growth of the cells	Check the quality of hPSCs.New line of hPSCs should be thawed.
34	Poor differentiation of hPSCs to SpM	Cell density is too high	Check plated cell density and cell confluency at each process.
			Check cell distribution in the wells.
			Cells should be evenly distributed.
	Poor induction of pFG-SpM and aFG-SpM	Cell density is too high	Check plated cell density and cell confluency at each process.
			Check cell distribution in the wells.
		Cells should be evenly distributed.
		RA might be degraded by light	Use fresh and photo-protected RA.
38	Cells lift off the plate	Cell density is too high	Check plated cell density and cell confluency at each process.
			Check cell distribution in the wells.
			Cells should be evenly distributed.
	Poor differentiation of hPSCs to organ-specific mesoderm	Cell density is too high	Check plated cell density and cell confluency at each process.
			Check cell distribution in the wells.
			Cells should be evenly distributed.