Fig. 4.

Serial spheroid formation capacity of colonospheres. To evaluate the self-renewal properties of generated spheroids, secondary spheroid formation was performed by serial passaging. Cells derived from a–f HT-29 and e–h Caco-2 spheroids generated spheroids for four consequence passage (P1–P4). There were no obvious differences in size and morphology of spheroids in different passages and their derived cells maintain their self-renewal capacity over several passages