Human Kidney Clonal Proliferation Disclose Lineage-restricted Precursor Characteristics

Overview

Journal Sci Rep

Specialty Science

Date 2020 Dec 17

PMID 33328501

Citations 2

Authors

Osnat Cohen-Zontag

Rotem Gershon

Orit Harari-Steinberg

Itamar Kanter

Dorit Omer

Oren Pleniceanu

Gal Tam

Sarit Oriel

Herzel Ben-Hur

Guy Katz

Zohar Dotan

Tomer Kalisky

Benjamin Dekel

Naomi Pode-Shakked

Affiliations

Soon will be listed here.

Abstract

In-vivo single cell clonal analysis in the adult mouse kidney has previously shown lineage-restricted clonal proliferation within varying nephron segments as a mechanism responsible for cell replacement and local regeneration. To analyze ex-vivo clonal growth, we now preformed limiting dilution to generate genuine clonal cultures from one single human renal epithelial cell, which can give rise to up to 3.4 * 10 cells, and analyzed their characteristics using transcriptomics. A comparison between clonal cultures revealed restriction to either proximal or distal kidney sub-lineages with distinct cellular and molecular characteristics; rapidly amplifying de-differentiated clones and a stably proliferating cuboidal epithelial-appearing clones, respectively. Furthermore, each showed distinct molecular features including cell-cycle, epithelial-mesenchymal transition, oxidative phosphorylation, BMP signaling pathway and cell surface markers. In addition, analysis of clonal versus bulk cultures show early clones to be more quiescent, with elevated expression of renal developmental genes and overall reduction in renal identity markers, but with an overlapping expression of nephron segment identifiers and multiple identity. Thus, ex-vivo clonal growth mimics the in-vivo situation displaying lineage-restricted precursor characteristics of mature renal cells. These data suggest that for reconstruction of varying renal lineages with human adult kidney based organoid technology and kidney regeneration ex-vivo, use of multiple heterogeneous precursors is warranted.

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