. Author manuscript; available in PMC: 2022 Aug 1.

Published in final edited form as: Nat Protoc. 2021 Jun 28;16(8):3874–3900. doi: 10.1038/s41596-021-00562-w

Table 7.

Guidelines to increase the success rate in usable monolayers

Steps	Good practices	Threshold
Organoid maintenance	High yield of organoids	Figure 4
	Appropriate size of organoids (>100 μm)	Figure 3, Figure 4
	Short incubation in CRS	≤60 min
	High cell viability diameter	≥80%
	Use of low centrifugal force for pelleting organoids	≤1000 g
Monolayer generation	Short incubation in CRS	≤60 min
	High cell viability	≥80%
	Sufficient cells for seeding	300,000
	Seeding of single cells	N/A
Monolayer differentiation	Confluent monolayer prior to differentiation	Figure 6b
Monolayer differentiation	Moderate TEER values (donor dependent) prior to differentiation	Donor dependent (>200 Ω cm²)