iPSC Kidney Organoid Differentiation Protocol

WTC11 induced pluripotent stem cell (iPSC) between passages 30 and 60 were used. Kidney organoid differentiation was induced as described previously [7 (link)]. In brief, iPSCs were plated at a density of 5,000 cells/well in a 24-well plate in mTeSR1 medium (Stem Cell Technologies) + 10 µM Y27632 (LC Laboratories) on plates (SPL Life Sciences) coated with 1% GelTrex (Thermo Fisher Scientific) (day –3). The medium was exchanged with 1.5% GelTrex in mTeSR1 (day –2), mTeSR1 (day –1), RPMI (Thermo Fisher Scientific) + 12 µM CHIR99021 (Tocris, Bristol, UK) (day 0), or RPMI + B27 supplement (Thermo Fisher Scientific) (day 1.5) and cells were fed every 2–3 days to promote kidney organoid differentiation. Organoids were analyzed on day 18.

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Park K., Lee J.Y., Lee S.Y., Jeong I., Park S.Y., Kim J.W., Nam S.A., Kim H.W., Kim Y.K, & Lee S. (2022). Deep learning predicts the differentiation of kidney organoids derived from human induced pluripotent stem cells. Kidney Research and Clinical Practice, 42(1), 75-85.

Publication 2022

mTeSR1 medium Y27632 GelTrex RPMI CHIR99021 B27 supplement

Cells Chir99021 Ipscs Kidney Organoids Stem cell Supplement Y27632

Corresponding Organization :

Other organizations : Pohang University of Science and Technology, Catholic University of Korea

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Variable analysis

independent variables

Passage number of iPSCs (between 30 and 60)
Plating density of iPSCs (5,000 cells/well)
Culture media used (mTeSR1, RPMI, B27 supplement)
Plate coating (1% GelTrex, 1.5% GelTrex)
CHIR99021 treatment (12 µM)

dependent variables

Kidney organoid differentiation

control variables

Tissue culture plates (SPL Life Sciences)
Y27632 treatment (10 µM)
Duration of differentiation (18 days)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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