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Self-renewal and differentiation of rat Epididymal basal cells using a novel in vitro organoid model

Pinel, Laurie et Cyr, Daniel G. ORCID: https://orcid.org/0000-0002-6566-783X (2021). Self-renewal and differentiation of rat Epididymal basal cells using a novel in vitro organoid model Biology of Reproduction , vol. 105 , nº 4. p. 987-1001. DOI: 10.1093/biolre/ioab113.

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Résumé

The epididymis is composed of a pseudostratified epithelium comprised of various cell types. Studies have shown that rat basal cells share common properties with adult stem cells and begin to differentiate in vitro in response to fibroblast growth factor and 5α-dihydrotestosterone. The characterization of rat basal cells is therefore necessary to fully understand the role of these cells. The objectives of this study were to assess the ability of single basal cells to develop organoids and to assess their ability to self-renew and differentiate in vitro. We isolated basal cells from the rat epididymis and established 3-dimensional cell cultures from the basal and non-basal cell fractions. Organoids were formed by single adult epididymal basal cells. Organoids were dissociated into single basal cells which were able to reform new organoids, and were maintained over 10 generations. Long-term culture of organoids revealed that these cells could differentiated into cells expressing the principal cell markers aquaporin 9 and cystic fibrosis transmembrane conductance regulator. Electron microscopy demonstrated that organoids were comprised of several polarized cell types displaying microvilli and the ability to form tight junctions. Additionally, organoids could be formed by basal cells from either the proximal or distal region of the epididymis, and are able to secrete clusterin, a protein implicated in the maturation of spermatozoa. These data indicate that rat basal cells can be used to derive epididymal organoids, and further supports that notion that these may represent a stem cell population in the epididymis.

Type de document: Article
Mots-clés libres: 3D Cell Culture; Basal Cells; Epididymis; Organoids; Stem Cells
Centre: Centre INRS-Institut Armand Frappier
Date de dépôt: 23 juin 2022 02:40
Dernière modification: 23 juin 2022 02:40
URI: https://espace.inrs.ca/id/eprint/12418

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