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. 2017 Dec 7;10(1):314–328. doi: 10.1016/j.stemcr.2017.11.004

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© 2017 The Author(s)

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

Characterization of Human iPSC-Derived Organoids Cultured in Happy Cell ASM

(A) Relative mRNA levels of the indicated genes in TkDN4-M-derived organoids cultured with Matrigel (open bars) or Happy Cell ASM (closed bars) plus 25% WRN CM and requisite ingredients for human organoid culture after one (P1) or two (P2) passages in each culture condition. The relative mRNA levels were determined by qPCR and normalized to GAPDH. The assays were performed in three independent biological replicates. Data presented as mean ± SEM. ^∗p < 0.05, Student's t test.

(B) Relative mRNA levels of the indicated genes in TkDN4-M-derived organoids cultured in Happy Cell ASM with or without 300 ng/mL recombinant mWNT3A for 6 days. The relative mRNA levels were determined by qPCR and normalized to GAPDH. The assays were performed in three independent biological replicates. Data presented as mean ± SEM. ^∗p < 0.05, Student's t test.

(C) Whole-mount images of TkDN4-M-derived organoids cultured in Happy Cell ASM with or without 300 ng/mL recombinant mWNT3A after staining with phalloidin (blue) and anti-MKI67 antibodies. Scale bars, 50 μm.