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. 2021 Feb 22;10:610368. doi: 10.3389/fcimb.2020.610368

Figure 1.

Figure 1

Characterization and differentiation of 3D spheroids/organoids from human, mouse, pig and chicken origin. (A) A new spheroid culture was cultured for 3 days (5 days for human) in WERN medium and then spheroids were either maintained in WERN medium or differentiated in ERN medium for further 3 days (human for 5 days). At indicated time points, spheroids/organoids were imaged and/or harvested for RNA extraction and transcriptional analysis by RT-qPCR. (B) Representative brightfield microscopic images of spheroids/organoids. Scale bars represent 500 µm (upper panel in each species) and 100 µm (lower panel in each species). (C) Quantification of marker genes specific for various intestinal cell types by RT-qPCR. Note that the data are normalized to the transcript abundance of 3/5 day WERN spheroid culture. EE, enteroendocrine cell; PC, Paneth cell; GC, goblet cell; SC, stem cell; E, enterocyte. RT-qPCR experiments show mean (± 95% CI) of ≥ 4 technical replicas of at least two independent biological replicates. n.d., not detectable.