University of Oulu

Rak-Raszewska, A, Reint, G, Geiger, F, Naillat, F, Vainio, SJ. Deciphering the minimal quantity of mouse primary cells to undergo nephrogenesis ex vivo. Developmental Dynamics. 2022; 251( 3): 536- 550. doi:10.1002/dvdy.418

Deciphering the minimal quantity of mouse primary cells to undergo nephrogenesis ex vivo

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Author: Rak-Raszewska, Aleksandra1; Reint, Ganna1; Geiger, Fabienne1;
Organizations: 1Laboratory of Developmental Biology, Disease Networks Researtch Unit, Faculty of Biochemistry and Molecular Medicine, University of Oulu, Oulu, Finland
2Kvantum Institute, Infotech Oulu, University of Oulu, Oulu, Finland
Format: article
Version: published version
Access: open
Online Access: PDF Full Text (PDF, 21.5 MB)
Persistent link: http://urn.fi/urn:nbn:fi-fe2022030722223
Language: English
Published: John Wiley & Sons, 2022
Publish Date: 2022-03-07
Description:

Abstract

Background: Tissue organoids derived from primary cells have high potential for studying organ development and diseases in numerous organs. They recreate the morphological structure and mimic the functions of given organ while being compact in size, easy to produce, and suitable for use in various experimental setups.

Results: In this study we established the number of cells that form mouse kidney rudiments at E11.5, and generated renal organoids of various sizes from the mouse primary cells of the metanephric mesenchyme (MM). We investigated the ability of renal organoids to undergo nephrogenesis upon Wnt/ β-catenin pathway—mediated tubule induction with a GSK-3 inhibitor (BIO) or by initiation through the ureteric bud (UB). We found that 5000 cells of MM cells are necessary to successfully form renal organoids with well-structured nephrons as judged by fluorescent microscopy, transmission electron microscopy (TEM), and quantitative Polymerase Chain Reaction (qPCR). These mouse organoids also recapitulated renal secretion function in the proximal tubules.

Conclusions: We show that a significant decrease of cells used to generate renal mouse organoids in a dissociation/re-aggregation assay, does not interfere with development, and goes toward 3Rs. This enables generation of more experimental samples with one mouse litter, limiting the number of animals used for studies.

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Series: Developmental dynamics
ISSN: 1058-8388
ISSN-E: 1097-0177
ISSN-L: 1058-8388
Volume: 251
Issue: 3
Pages: 536 - 550
DOI: 10.1002/dvdy.418
OADOI: https://oadoi.org/10.1002/dvdy.418
Type of Publication: A1 Journal article – refereed
Field of Science: 3111 Biomedicine
Subjects:
3Rs
Funding: This work was supported by Finnish Cultural Foundation (#00160821 personal grant to Aleksandra Rak-Raszewsko), CIMO Fellowship (personal grant to Ganna Reint), Center of Excellence grant (#2513149) and Academy of Finland Grant (315030).
Academy of Finland Grant Number: 315030
Detailed Information: 315030 (Academy of Finland Funding decision)
Copyright information: © 2021 The Authors.This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
  https://creativecommons.org/licenses/by-nc-nd/4.0/