Journal: Kidney360

Article Title: A Novel Human Distal Tubuloid-on-a-Chip Model for Investigating Sodium and Water Transport Mechanisms

doi: 10.34067/KID.0000000992

Figure Lengend Snippet: Development and characterization of tubuloid-on-chip model. (A) Experimental timeline and platform schematic. Timeline shows cell seeding (Day 0), followed by growth period (Day 5) and subsequent functional analysis (Day 5+7). The OrganoPlate system features three compartments: two perfusable channels flanking a central ECM gel chamber, enabling tubuloid formation and culture. (B) Brightfield microscopy showing progressive tubule development: initial cell seeding (Day 0), early attachment (Day 2), and mature tubule formation with visible doming (Day 5+7). Asterisks (*) indicate the formation of subepithelial domes (representative images, n =64 chips). (C) Immunofluorescence 3D reconstruction demonstrating tubule polarization and barrier formation. (D) Confocal projections of a tubuloid chip in the 3-lane microfluidic chip stained for ZO-1 (green), Na + /K + -ATPase (red), and nuclei (blue). Panels display XY, YZ, and XZ projections, illustrating the epithelial monolayer's polarization and spatial organization. The right panel shows a magnified view with a cross-section highlighting the polarized epithelial layer. 3D, three-dimensional; ECM, extracellular matrix; qPCR, quantitative PCR.

Article Snippet: We developed a human distal tubuloid-on-a-chip model by integrating primary human tubuloid cells with the OrganoPlate microfluidic culture system.

Techniques: Functional Assay, Microscopy, Immunofluorescence, Staining, Real-time Polymerase Chain Reaction

Journal: Kidney360

Article Title: A Novel Human Distal Tubuloid-on-a-Chip Model for Investigating Sodium and Water Transport Mechanisms

doi: 10.34067/KID.0000000992

Figure Lengend Snippet: Expression profile of distal nephron markers in differentiated tubuloids. (A) Left: Schematic illustration of DNS highlighting the TAL of the loop of Henle (TAL; SLC12A1/NKCC2) and CD (SCNN1a/ENaC, AQP2) markers. Right: qPCR analysis showing segment-specific gene expression across four donors after differentiation (fold change relative to basal medium, log scale, Mean±SD). Samples from five chips per donor were pooled to create each replicate, with three independent samples per condition for each of the four donors. (B) Immunofluorescence analysis of tubuloid cultures (20× magnification). Upper panels show full-length tubules; lower panels show magnified regions (dotted boxes). Staining shows expression of ZO-1 (tight junctions), ECAD (distal tubule), NKCC2 (TAL), and AQP2 (CD), all counterstained with DNA (representative images, n =2–3). (C) Comparative gene expression analysis between 3D (OrganoPlate) and 2D (tubuloids grown on static 24-well plates) culture conditions across four donors (fold change, n =3). Comparisons made within each donor due to variability in baseline expression. All qPCR data were normalized to the housekeeping gene GAPDH and expressed as fold change relative to the corresponding 2D static culture condition for each donor. 2D, two-dimensional; CD, collecting duct; DCT, Distal convoluted tubule; DNS, distal nephron segments; ECAD, E-cadherin; ENaC, epithelial sodium channel; PT, proximal tubule; TAL, thick ascending limb.

Article Snippet: We developed a human distal tubuloid-on-a-chip model by integrating primary human tubuloid cells with the OrganoPlate microfluidic culture system.

Techniques: Expressing, Gene Expression, Immunofluorescence, Staining