Department of Surgery, School of Medicine, Virginia Commonwealth University, Richmond, VA, USA.
Hume-Lee Transplant Center, VCU Health System, Richmond, VA, USA.
Cell Transplant. 2022 Jan-Dec;31:9636897221086966. doi: 10.1177/09636897221086966.
Islet culture before clinical transplantation has been adopted by various centers, but its effect on the survival and function of islets relative to the culture conditions and media needs further assessment. Human islets were cultured or preserved under four different conditions and three media options. Parameters such as recovery, viability, function, islet damage, and gene expressions for markers of hypoxia, and inflammation were assessed after 48-h culture or preservation. Preservation of islets was performed at 4°C in Connaught's Medical Research Lab (CMRL) and University of Wisconsin (UW) media. Islets were cultured at 22°C, 37°C, and 37°C-22°C in CMRL and PRODO culture media. Islets preserved in UW solution had visually good morphology and exhibited higher recovery with less islet damage compared with the rest of the groups, whereas islets preserved in CMRL at 4°C resulted in poor morphology, recovery, viability, and function compared with the rest of the treatment conditions. Culture at 22°C and 37°C demonstrated an increase in the expression of inflammatory and hypoxia-related genes. In conclusion, islets preserved at 4°C in UW solution showed the best overall outcomes after 48 h compared with islets cultured at 22°C, 37°C, or 37°C-22°C in PRODO. Advancement in islet culture media is warranted to reduce inflammatory gene activation and improve recovery of islets for transplantation.
在临床移植前，胰岛培养已被各个中心采用，但它对胰岛存活和功能的影响相对于培养条件和培养基的选择仍需进一步评估。本研究将人类胰岛在四种不同条件和三种培养基选择下进行培养或保存。在 48 小时培养或保存后，评估胰岛的回收、活力、功能、胰岛损伤以及缺氧和炎症标志物的基因表达等参数。胰岛在 Connaught 医学研究实验室（CMRL）和威斯康星大学（UW）培养基中于 4°C 下保存。胰岛在 22°C、37°C 和 37°C-22°C 下于 CMRL 和 PRODO 培养基中进行培养。与其他组相比，UW 溶液保存的胰岛具有良好的形态，且胰岛损伤较少，表现出更高的回收率，而在 4°C 下的 CMRL 中保存的胰岛与其他处理条件相比，形态、回收率、活力和功能较差。在 22°C 和 37°C 下培养显示出炎症和缺氧相关基因表达增加。总之，与在 22°C、37°C 或 37°C-22°C 下于 PRODO 中培养相比，UW 溶液中 4°C 保存的胰岛在 48 小时后显示出最佳的总体结果。需要改进胰岛培养基，以减少炎症基因的激活，并提高胰岛移植的回收率。
