Dual-Lineage Kidney Models for Epithelial Transport and Linked Drug-Injury Comparisons

Research Applications

Compare segment formation, renal epithelial uptake/efflux and drug injury with or without linked intestinal/hepatic components.

Model & Design

H9 human embryonic stem cells and three tested iPSC sources (fibroblast/PBMC/cord blood). Caco-2 intestinal and HepaRG hepatic cell lines support the separate circuit arm.

Experimental Format

Transient 0.1 μM retinoic acid during early mesoderm induction; BMP7 at 100 ng/mL plus FGF9 maturation 10–14 days. Organoid-derived renal epithelial monolayers (KORTECs) at air–liquid interface for transport; circulating full network versus kidney-alone comparison.

Model Components

• Component

  Functional controls

  Description

  HK-2 cell-line and empty-insert barriers, albumin uptake and CDFDA-derived CDF efflux with or without MRP2/4 inhibitor MK571. Alpelisib was detected after 48 hours, but the narrative and figure caption report opposite application/transfer directions.

• Component

  Drug injury

  Description

  The main Results report apoptosis after 25 μM cisplatin, 20 μM cyclosporine A and 2.5 μM doxorubicin, plus gentamicin-associated segment injury. Circuit acetaminophen concentration, exposure duration and flow conditions are not specified in the main methods.

Measurements & Endpoints

• Segment markers, polarity/cilia and developmental transcriptomic match
• TEER, dextran leakage and albumin uptake
• Inhibitor-sensitive fluorescent efflux
• Drug-associated apoptosis/injury markers
• APAP–GSH, APAP–cysteine and 4-aminophenol in circuit medium

Analysis

Differentiation, derived-monolayer transport and whole-organoid toxicity are separate experimental arms. The full Caco-2/HepaRG/kidney circuit is compared with kidney alone in the same apparatus. Measured metabolite changes accompany greater renal injury; individual-metabolite causality and NAPQI concentration were not established.

Published Evidence

• Study Design

  RA/BMP perturbation, developmental reference comparison, HK-2/barrier/efflux controls and linked-culture drug injury.

  Study Sample

  6,476 day 18 organoid cells for single-cell profiling; three iPSC sources tested alongside H9. Circuit injury-marker n = 4; segment-caspase n = 6.

  Comparison

  No/low/high or prolonged RA, FGF9 only versus BMP7, signaling inhibitors, HK-2 and MK571 controls, full network versus kidney-alone.

  Finding

  Brief low-dose RA and BMP7 improved segment/polarity markers. KORTECs had stronger barrier/albumin uptake than HK-2; MK571 reduced apical CDF transfer. Full network increased APAP-associated injury alongside measured metabolites compared with kidney-alone circuit.

  Interpretation & Limits

  • Developmental reference comparisons used fetal and newborn kidney; adult filtration or urine production was not validated.
  • Inconsistent alpelisib transfer directions and the absence of a dedicated mechanistic test limit drug-specific transport interpretation.
  • The main text omits the circuit APAP dose and exposure period and the direct-nephrotoxicant exposure duration, limiting reproduction of those arms.
  • Integrated native vascular and immune compartments and patient-response prediction were not established.