Deciphering the minimal quantity of mouse primary cells to undergo nephrogenesis ex vivo. Issue 3 (16th September 2021)

• Record Type: Journal Article
• Authors: Rak‐Raszewska, Aleksandra; Reint, Ganna; Geiger, Fabienne; Naillat, Florence; Vainio, Seppo J.
• Is Part Of: Developmental dynamics. Volume 251:Issue 3(2022)
• Page Start: 536
• Page End: 550
• Publication Date: 2021-09-16
• Subjects: 3Rs -- dissociation/re‐aggregation assay -- kidney development -- minimal cell number -- renal organoids; Morphogenesis -- Periodicals; Anatomy -- Periodicals; Anatomie -- Périodiques; Biologie du développement -- Périodiques; 571.833
• DOI: 10.1002/dvdy.418
• Languages: English
• ISSNs: 1058-8388

Abstract

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. Key Findings: Generation of functional renal mouse organoids requires to reach a minimal cell number. The type of cells generating renal organoids is also important. Reduction of cells required for generation of organoids reduce the number of animals used and goes well with 3R's.