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Background:

The distal nephron segments are essential for electrolyte, acid-base and volume homeostasis. Tubuloid culture allows ample expansion (exp) of primary human renal epithelium from urine or tissue. Here we differentiate tubuloids into thick ascending limb of Henle (TAL) and principal cells (PC), characterize these at mRNA, protein and functional levels, and establish an in vitro human lithium (Li+) tubulopathy model.

Methods:

Tubuloids were grown from kidney tissue of 4 donors in exp medium. After differentiation (diff) for 7 days, tubuloids were studied by qPCR, single cell RNA sequencing (scSeq) and immunofluorescence (IF)/-histochemistry (IHC). ScSeq data was compared with human kidney tissue. Electrolyte transport was tested using the FluxOR™ II assay. Diff tubuloids were exposed to 10 mM Li+ or Na+(control) and analyzed by the same techniques.

Results:

Diff increased transcription of NKCC2 …