Table 2.

Recommended number of cells per cryotube and number of wells to thaw a cryotube into for hPSCs, CPCs, EpiCs, and EpiC-FBs

Cell type	Suggested number of wells per cryotube	Approximate cell count per cryotube	Number of wells to thaw cryotube of cells into	Medium to thaw cells into	Maintenance medium
hPSCs	1 confluent well from a 6 well plate	1–2 M	3–6 wells of a 6 well plate, will be confluent in 2–4 days depending on hPSC line, hPSC passage number, etc.	mTeSR1 + 5 mM Y-27632	mTeSR1
CPCs	4–6 wells from 12 well plate	6–12 M	4–6 6 well plates (for EpiC differentiation)	LaSR + 5 mM Y-27632	Directly differentiate into EpiCs, cannot be maintained as CPCs
EpiCs	Entire confluent 6 well plate	1 M	1 6 well plate ∗generally notice a lot of cell death, and observed better survival if replated at a higher density, will be confluent in 3–6 days	LaSR + 5 mM Y-27632 + 0.5 μM A83-01	LaSR + 0.5 μM A83-01
EpiC-FBs	1 confluent well from a 6 well plate	1 M	1–2 6 well plates, Will be confluent in 4–7 days	FibroGRO	FibroGRO