Endothélialisation d'un modèle 3D de muqueuse vaginale humaine reconstruite par génie tissulaire : modélisation in vitro et implantation animale
|Abstract:||Tissue engineering of autologous human vaginal mucosa (HVM) introduces novel surgical applications to the field of vaginal reconstruction for paediatric patients with congenital urogenital abnormalities such as the Mayer-Rokitansky-Küster-Hauser syndrome (MRKH) or neoplastic diseases. Vascularization of tissue-engineered constructs represents a major challenge seeing that graft survival and success rate highly depend on it. This study aims at reconstructing a pseudo-capillary network within a tissue-engineered HVM using the self-assembly technique free of exogenous materials. Vaginal stromal cells were co-seeded with endothelial cells derived from a human umbilical cord vein (HUVEC). Different cell culture techniques were tested, the classical self-assembly (SA) by sheet stacking, re-seeding (RS) and a new hybrid SA/RS method in order to determine the approach that is the most adapted for pre-vascularization of the HVM, while maintaining biomechanical properties that are suitable for surgery. The presence of a pseudo-capillary network in vitro was assessed with specific markers such as PECAM-1/CD31 and Von Willebrand factor. Additionally, neuroglial antigen 2 was detected at the periphery of capillaries and reveals the presence of pericytes within constructs produced with the RS and SA/RS methods. Our results show that the use of a combined SA/RS technique seems to be most adapted for the pre-vascularization of the HVM as it generates constructs with higher microvascular maturity and mechanical properties compatible with surgical handling. Transduced HUVEC with a vector that allows the expression of GFP and luciferase were used to observe the formation of a capillary network in vitro and to monitor endothelial cells in vivo. To assess the functionality of the reconstructed capillary-like network, endothelialized HVM constructs were implanted in immunocompromized mice. The finding of mouse red blood cells within GFP positive capillaries confirms the functionality of the reconstructed capillary-like network in vivo. Finally, this first tissue-engineered endothelialized HVM model can be used for numerous clinical and research applications.|
|Document Type:||Mémoire de maîtrise|
|Open Access Date:||24 April 2018|
|Collection:||Thèses et mémoires|
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