Table 1.

Comparison of the results obtained for 3D spheroids with the three different methods.

	3D culture methods
	96-well hanging drop plate	24-well plate with a cell-repellent surface	Ultra-low attachment 96-well plates
Cell seeding	Easy	Easy	Easy
Easiness in spheroids formation	Easy	Intermediate	Easy
Morphology	Round type with jagged edges	Round-type	Round-type
Culture maintenance	Difficult	Difficult	Easy
Medium change	Difficult	Difficult	Difficult
Spheroids monitoring	Difficult	Intermediate	Easy
Picture quality	Low	High	High
Spheroid perimeter measurement reproducibility error	2.9 × 106 pixels	N. A.	1.2 × 106 pixels
Possibility to perform assays into the seeding plate	No	No	Yes
Overall time of the experiment	7 days	10 days	7 days

Reproducibility error is not available for spheroids seeded with the second method since several spheroids were thrown away during medium refreshing.