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Ingénierie tissulaire de l'urothélium

Abstract : The urothelium is a specialized epithelium covering the urinary excretory system. In case of lesions, it is replaced by segments of digestive tract. An alternative could be the creation of an in vitro neotissue covered with urothelium. The objective is to evaluate the potential of different cell types to develop an in vitro urothelium: 1) a novel source of human urothelial cells (UC), 2) urine-derived stem cells (UDSC) and 3) Wharton's jelly-derived mesenchymal stem cells (WJ-MSC). Material and method: UC are obtained by digestion of bladder neck biopsies during Trans Urethral Prostate Resection (TURP). UDSCs are obtained by centrifugation of urine. WJ-MSCs are obtained by culture of Wharton's jelly fragments. Expansion of UC is performed in keratinocyte culture medium. Stem cell expansion includes an amplification phase (maintenance of proliferation), followed by a uroinduction phase (development of urothelial characteristics). The cells are then seeded on a bovine collagen biomaterial and cultured in 3D for 28 days. Histological, immunohistological and molecular biology (RT-qPCR) analyses were performed. Results: TURP biopsies show a damaged urothelium, a small contingent of UC is extracted, adhering and proliferating with difficulty and dedifferentiation. The 3D culture induces a heterogeneous colonization by cells not expressing cytokeratins. UDSCs are easily obtained, have a rapid proliferation without dedifferentiation and allow a regular cellularization by cytokeratin + cells. WJ-MSCs strongly express strain markers, the uro-induction phase has a heterogeneous effect, cellularized biomaterials show a thin layer of cells with sometimes a label for cytokeratin. Conclusion: TURP biopsies allow to routinely obtain human UCs that are not very usable in tissue engineering because of expansion and adhesion difficulties. UDSCs are easily obtained non-invasively and allow to obtain biomaterials cellularized by cells expressing urothelial markers. WJ-MSCs are incompletely oriented to a urothelial phenotype by uroinduction, but can colonize the biomaterial under good conditions.
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https://hal.univ-lorraine.fr/tel-03417112
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Submitted on : Friday, November 5, 2021 - 3:12:06 PM
Last modification on : Saturday, November 6, 2021 - 4:20:37 AM

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  • HAL Id : tel-03417112, version 1

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Nicolas Berte. Ingénierie tissulaire de l'urothélium. Médecine humaine et pathologie. Université de Lorraine, 2021. Français. ⟨NNT : 2021LORR0138⟩. ⟨tel-03417112⟩

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