Fig. 3: Generation of mature kidney organoids in UniMat. | Nature Communications

Fig. 3: Generation of mature kidney organoids in UniMat.

From: Scalable production of uniform and mature organoids in a 3D geometrically-engineered permeable membrane

Fig. 3: Generation of mature kidney organoids in UniMat.The alternative text for this image may have been generated using AI.

a Confocal images of kidney organoids in AggreWell and UniMat, stained with PODXL, LTL, and CDH1. Scale bar = 50 µm. b Quantification of PODXL+, LTL+, and CDH1+ fractions in individual organoids in AggreWell and UniMat (n = 15 organoids, mean ± SD). Significance by one-way ANOVA with Tukey’s multiple comparisons test: P = 3.3Ă—10−11 (PODXL+); P = 2.4 × 10−8 (LTL+); P = 1.2 × 10−5 (CDH1+). c Immunofluorescence images of PODXL and ZO-1 of kidney organoids in AggreWell and UniMat. Scale bar = 20 µm. d Immunofluorescence images of LTL of kidney organoids in AggreWell and UniMat. Scale bar = 20 µm. Bottom plots show LTL intensity across line scans marked by red arrows in the images above. e Dextran uptake assay in LTL+ proximal tubules of kidney organoids in AggreWell and UniMat. Scale bar = 10 µm. f SEM images of microwell in UniMat-ZP, UniMat-LP, and original UniMat with magnified SEM images of microwell walls. Scale bar = 100 µm and 2 µm (Inset images). g Permeability coefficients across UniMat-ZP, UniMat-LP, and UniMat (n = 3 samples, mean ± SD). Significance by one-way ANOVA with Tukey’s multiple comparisons test: PUniMat-ZP, UniMat-LP = 7.5Ă—10−10, PUniMat-LP, UniMat = 1.1 × 10−8 (0.4 kDa); PUniMat-ZP, UniMat-LP = 1.4 × 10−7, PUniMat-LP, UniMat = 4.8 × 10−9 (3 kDa); PUniMat-ZP, UniMat-LP = 2.2 × 10−6, PUniMat-LP, UniMat = 9.1 × 10−11 (20 kDa); PUniMat-ZP, UniMat-LP = 3.6 × 10−5, PUniMat-LP, UniMat = 7.8 × 10−9 (40 kDa). h Numerical simulations of glucose concentration distribution around organoid in UniMat-ZP, UniMat-LP, and UniMat. i qRT-PCR of kidney organoids in UniMat-ZP, UniMat-LP, and UniMat (n = 5 independent experiments, mean ± SD). Significance by one-way ANOVA with Tukey’s multiple comparisons test: PUniMat-ZP, UniMat-LP = 0.0037, PUniMat-LP, UniMat = 8.2 × 10−8 (NPHS1); PUniMat-ZP, UniMat-LP = 1.0Ă—10−7, PUniMat-LP, UniMat = 3.1 × 10−10 (PODXL); PUniMat-ZP, UniMat-LP = 1.9 × 10−5, PUniMat-LP, UniMat = 0.0002 (ABCB1); PUniMat-LP, UniMat = 1.6 × 10−5 (AQP1); PUniMat-ZP, UniMat-LP = 0.0031, PUniMat-LP, UniMat = 1.4 × 10−5 (SLC34A1); PUniMat-ZP, UniMat-LP = 0.0104, (UMOD); PUniMat-ZP, UniMat-LP = 2.2 × 10−9 (CDH1). Source data are provided as a Source Data file.

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