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. Author manuscript; available in PMC: 2021 May 23.
Published in final edited form as: Nat Mater. 2020 Nov 23;20(2):260–271. doi: 10.1038/s41563-020-00853-9

Figure 6. Generation of a kidney tissue patch using 3D extrusion cellular bioprinting.

Figure 6.

A. Illustration of the scripted movement of the needle tip for cell paste extrusion, generating a patch organoid across an area of approximately 4.8mm × 6mm, containing approximately 4×105 cells. Lines indicate continuous movements. B. Brightfield imaging of the bioprinted kidney tissue patch demonstrating uniform formation of nephron structures. C. Live confocal imaging of MAFBmTAGBFP2 reporter signal throughout a patch organoid at D7+12. Scale bar =1mm. Image is representative of n = 3 replicate patches. D. Confocal immunofluorescence of a D7+14 patch organoid demonstrating uniform distribution of nephrons expressing markers for podocytes (mTagBFP2 [left panel; blue]), proximal tubules (LTL [left panel; green] and HNF4A [right panel; red]), nephron epithelium (EPCAM [left panel; red]), distal tubule/loop of Henle TAL (SLC12A1 [right panel; green]) and endothelial cells (SOX17 [right panel; grey]). Scale bars = 100μm. Representative image from n = 3 patches. E. Live confocal imaging of a D7+14 patch organoid derived from the HNF4AYFP reporter iPSC line following incubation in TRITC-albumin substrate. Images depict TRITC-albumin (red) uptake into YFP-positive proximal tubules (yellow). Outlined areas (whole organoid images) are shown at higher magnification below, with and without phase contrast overlays. Scale bars = 100μm. Assay performed on a representative sample from n = 3 patches.