Lebreton Fanny, Bellofatto Kevin, Wassmer Charles H, Perez Lisa, Lavallard Vanessa, Parnaud Géraldine, Cottet-Dumoulin David, Kerr-Conte Julie, Pattou François, Bosco Domenico, Othenin-Girard Véronique, Martinez de Tejada Begoña, Berishvili Ekaterine
Cell Isolation and Transplantation Center, Department of Surgery, Faculty Diabetes Center, Geneva University Hospitals and University of Geneva, Geneva, Switzerland.
INSERM U1190, Translational Research for Diabetes, University of Lille, France.
Am J Transplant. 2020 Jun;20(6):1551-1561. doi: 10.1111/ajt.15812. Epub 2020 Mar 6.
Hypoxia is a major cause of considerable islet loss during the early posttransplant period. Here, we investigate whether shielding islets with human amniotic epithelial cells (hAECs), which possess anti-inflammatory and regenerative properties, improves islet engraftment and survival. Shielded islets were generated on agarose microwells by mixing rat islets (RIs) or human islets (HI) and hAECs (100 hAECs/IEQ). Islet secretory function and viability were assessed after culture in hypoxia (1% O ) or normoxia (21% O ) in vitro. In vivo function was evaluated after transplant under the kidney capsule of diabetic immunodeficient mice. Graft morphology and vascularization were evaluated by immunohistochemistry. Both shielded RIs and HIs show higher viability and increased glucose-stimulated insulin secretion after exposure to hypoxia in vitro compared with control islets. Transplant of shielded islets results in considerably earlier normoglycemia and vascularization, an enhanced glucose tolerance, and a higher β cell mass. Our results show that hAECs have a clear cytoprotective effect against hypoxic damages in vitro. This strategy improves β cell mass engraftment and islet revascularization, leading to an improved capacity of islets to reverse hyperglycemia, and could be rapidly applicable in the clinical situation seeing that the modification to HIs are minor.
缺氧是移植后早期相当一部分胰岛丧失的主要原因。在此，我们研究用具有抗炎和再生特性的人羊膜上皮细胞（hAECs）保护胰岛是否能改善胰岛植入和存活。通过将大鼠胰岛（RIs）或人胰岛（HI）与hAECs（100个hAECs/胰岛当量）混合，在琼脂糖微孔上生成受保护的胰岛。在体外缺氧（1%O₂）或常氧（21%O₂）条件下培养后，评估胰岛分泌功能和活力。在糖尿病免疫缺陷小鼠肾被膜下移植后，评估体内功能。通过免疫组织化学评估移植物形态和血管化。与对照胰岛相比，在体外暴露于缺氧后，受保护的RIs和HIs均显示出更高的活力和增加的葡萄糖刺激胰岛素分泌。移植受保护的胰岛可导致更早出现正常血糖和血管化、增强的葡萄糖耐量以及更高的β细胞量。我们的结果表明，hAECs在体外对缺氧损伤具有明显的细胞保护作用。这种策略可改善β细胞量植入和胰岛再血管化，从而提高胰岛逆转高血糖的能力，并且鉴于对HIs的修饰较小，可迅速应用于临床情况。
