Concentrated medium collected from WEHI-YH2 cells (cYH2CM) stimulates colon crypt formation.

<p>Conditioned Medium (CM) from WEHI-YH2 cells (YH2CM) was collected from confluent cultures incubated for different times (24, 48 and 72 hours) in DMEM/F12 with the N2 and B27 supplements. The CM was concentrated 10-fold using a Centriprep YM3 filter (with a 3kDa molecular weight cut-off) (cYH2CM). Different amounts of YH2CM and cYH2CM were used to grow colon crypts in culture and the results are compared with the WEHI-YH2 colon crypt co-cultures. Images of the cultures were taken on Day 0 and Day 6, processed and scored for either colonospheres or organoids with crypts: (A) colonosphere- and (B) colon crypt–forming efficiency on Day 6 (the number of colonospheres or colonoids on Day 6 divided by the number of crypts plated on Day 0). The dotted lines show the colonosphere and colon colonoid formation efficiency in co-cultures with WEHI-YH2 cells. Error bars: SEM, n = 4 from 2 independent experiments of duplicates, 2-way ANOVA multiple comparisons Tukey’s test.