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. 2022 Feb 15;13:848433. doi: 10.3389/fimmu.2022.848433

Figure 2.

Figure 2

In vivo differentiation of chimeric renal organoids into renal tissue. (A) Images of transplanted chimeric renal organoids made from mouse and rat fetal kidneys under the renal capsule of immunodeficient mice (scale bars 2 mm). (B) Brightfield and fluorescence images of chimeric renal organoids 14 days after transplantation (scale bars, upper column: 1 mm, lower column: 1 mm). Recipient-derived blood vessels invade the organoid (yellow arrowhead). (C) Fluorescence microscopy and HE staining images of frozen sections (scale bar 500 μm). (D) Fluorescence immunostaining image of a frozen section of the whole renal organoid (scale bar 200 μm). Multiple Ecad-positive tubular structures. (E) Nephrin-positive glomeruli and PDGFRb-positive renal stroma in the organoid (scale bar 200 μm). (F) GATA3-positive and Ecad-positive collecting ducts consisted of GFP-positive rat cells and GFP-negative mice cells (scale bar 20 μm). (G) Aquaporin 2-positive and GATA3-positive principal cells consisted of GFP-positive rat cells and GFP-negative mice cells (scale bar 20 μm). (H) V-ATPaseB1-positive and GATA3-positive intercalated cells consisted of GFP-positive rat cells and GFP-negative mice cells (scale bar 50 μm). (I) Connection of GFP-negative and CK8-positive mouse collecting ducts to GFP-positive and Ecad-positive rat distal tubules (scale bar 20 μm). (J) NKCC2-positive loop of Henle consisting of GFP-positive rat cells and GFP-negative mice cells (scale bar 20 μm). (K) GFP-positive and megalin-positive proximal tubules connected to the tubules of GFP-negative mice cells (scale bar 50 μm). (L) Nephrin-positive podocytes and PDGFRb-positive mesangial cells generated from GFP-positive rat cells and GFP-negative mice cells in the glomeruli (scale bar 20 μm). (M) PDGFRb-positive interstitial fibroblasts consisted of GFP-positive rat cells and GFP-negative mice cells (scale bar 20 μm). (N) α SMA-positive vascular pericytes from GFP-positive rat cells (scale bar 10 μm). (O) Renin-producing cells were found around the afferent arterioles of the glomeruli (scale bar 20 µm). (P) Contribution of GFP-positive rat cells to the three lineages of collecting ducts, nephrons, and stroma. The percentages of GFP-positive cells in GATA3-positive collecting ducts, Ecad-positive, and GATA3-negative nephron tubules, and PDGFRb-positive stroma are shown. Error bars in the graphs represent the standard error of the mean (each group, n = 6).