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Feeder cells were seeded with 1.25 10 4 cells/well in a 96-well plate and cultured for 1 day in the medium including bFGF and Y-27632, and iPS cells were additionally seeded with 5000 cells/well and cultured for 1 day
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The ABX, sucrose, and selective antibiotic water were then filtered through the 0.22 m filter (500 ml, Sorfa) for sterilization
Subsequently, organoids were incubated for 2h in HEPES-saline buffer supplemented with either LPS (10g/mL), MDP (10g/mL), a combination of TNF (10ng/mL) and IFN (50 ng/mL), DCA (30 M) (as physiological positive control), or a mixture of 10 M Forskolin and 10 M 3-isobutyl-1-methylxanthine (F/I, for maximal hormone output)
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