Fig. 3

Fgf7 but Fgf10 induces differentiation of organoid cells. (A) Experimental protocol. AdBOs were grown for the first week (Day 1 to 8) in Fgf7+/Fgf10 + medium, split to multiple wells, and divided into 5 groups. Each group of organoids was cultured in Fgf7+/Fgf10+, Fgf7+/Fgf10-, Fgf7-/Fgf10+, 4xFgf7+/Fgf10-, or Fgf7-/4xFgf10 + medium for additional 1 week (Day 8 to 15) and then used for the following assays as indicated. (B) and (C) Relative expression levels of differentiated cell marker genes (Krt20, Upk1a, Upk2, Upk3a) (B) and immature cell marker genes (Krt5, Trp63, Krt14) (C) in organoids cultured in Fgf7+/Fgf10+, Fgf7+/Fgf10-, or Fgf7-/Fgf10 + medium. ***p < 0.001, **p < 0.01 and *p < 0.05. (D) AdBOs were grown for 1 week (Day 1 to 8) in Fgf7+/Fgf10 + medium and then cultured under 5 different conditions for additional 1 week (Day 8 to 15). Representative images of organoids in each group acquired on Day 15 were shown. Scale bar: 500 μm. (E) The number of viable cells in AdBOs cultured under 5 different conditions was assessed on Day 15 by Trypan blue exclusion assay. Values represent the average of three independent experiments and error bars denote standard error of the mean. ***p < 0.001, **p < 0.01 and *p < 0.05