Pancreatic islet cryopreservation by vitrification achieves high viability, function, recovery and clinical scalability for transplantation.
Pancreatic islet cryopreservation by vitrification achieves high viability, function, recovery and clinical scalability for transplantation.
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DOI：
10.1038/s41591-022-01718-1
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发表时间：
2022-04
期刊：
影响因子：
82.9
通讯作者：
Finger, Erik B.
中科院分区：
文献类型：
--
作者：
Zhan, Li;Rao, Joseph Sushil;Sethia, Nikhil;Slama, Michael Q.;Han, Zonghu;Tobolt, Diane;Etheridge, Michael;Peterson, Quinn P.;Dutcher, Cari S.;Bischof, John C.;Finger, Erik B.
胰岛移植可以治愈糖尿病，但需要足够数量的可获得的高质量胰岛。冷冻保存可以解决胰岛供应链的挑战，使质量控制的银行和捐赠胰岛池。不幸的是，冷冻保存在这一目标上没有成功，因为它必须同时提供高回收率、活力、功能和可扩展性。在这里，我们通过全面优化冷冻保护剂（CPA）组成、CPA装载和卸载条件以及玻璃化和复温（VR）方法，在小鼠、猪、人和人干细胞（SC）衍生的β细胞（SC-β）胰岛中实现了这一目标。相对于对照组，VR后的胰岛存活率，小鼠为90.5%，SC-β为92.1%，猪为87.2%，人为87.4%，并且在低温储存至少9个月后保持不变。VR胰岛具有正常的宏观、微观和超微结构形态。线粒体膜电位和三磷酸腺苷（ATP）水平轻微降低，但细胞呼吸的所有其他指标（包括产生ATP的耗氧率（OCR））均未发生变化。VR胰岛在体外和体内均具有正常的葡萄糖刺激胰岛素分泌（GSIS）功能。猪和SC-β胰岛在异种移植模型中产生胰岛素，在边缘团块同基因移植模型中检测的小鼠胰岛在移植后24-48小时内治愈了92%的受体的糖尿病。血糖控制良好，持续150天。最后，我们的方法处理了2，500个胰岛，在>89%的解冻后活力下具有>95%的胰岛回收率，并且可以容易地扩大规模以获得更高的通量。这些结果表明，冷冻保存现在可以用来提供所需的胰岛，以改善移植结果，治愈糖尿病。玻璃化冷冻方法的优化大大改善了胰岛的冷冻保存，并提高了糖尿病患者的移植结果。
Pancreatic islet transplantation can cure diabetes but requires accessible, high-quality islets in sufficient quantities. Cryopreservation could solve islet supply chain challenges by enabling quality-controlled banking and pooling of donor islets. Unfortunately, cryopreservation has not succeeded in this objective, as it must simultaneously provide high recovery, viability, function and scalability. Here, we achieve this goal in mouse, porcine, human and human stem cell (SC)-derived beta cell (SC-beta) islets by comprehensive optimization of cryoprotectant agent (CPA) composition, CPA loading and unloading conditions and methods for vitrification and rewarming (VR). Post-VR islet viability, relative to control, was 90.5% for mouse, 92.1% for SC-beta, 87.2% for porcine and 87.4% for human islets, and it remained unchanged for at least 9 months of cryogenic storage. VR islets had normal macroscopic, microscopic, and ultrastructural morphology. Mitochondrial membrane potential and adenosine triphosphate (ATP) levels were slightly reduced, but all other measures of cellular respiration, including oxygen consumption rate (OCR) to produce ATP, were unchanged. VR islets had normal glucose-stimulated insulin secretion (GSIS) function in vitro and in vivo. Porcine and SC-beta islets made insulin in xenotransplant models, and mouse islets tested in a marginal mass syngeneic transplant model cured diabetes in 92% of recipients within 24–48 h after transplant. Excellent glycemic control was seen for 150 days. Finally, our approach processed 2,500 islets with >95% islets recovery at >89% post-thaw viability and can readily be scaled up for higher throughput. These results suggest that cryopreservation can now be used to supply needed islets for improved transplantation outcomes that cure diabetes. Optimization of vitrification approaches substantially improves pancreatic islet cryopreservation for banking and boosts transplantation outcomes in diabetes.
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DOI：
10.4049/jimmunol.0803543
发表时间：
2009-03-01
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Melli K;Friedman RS;Martin AE;Finger EB;Miao G;Szot GL;Krummel MF;Tang Q
通讯作者：
Tang Q
影响因子：
0.9
作者：
Anazawa, T.;Balamurugan, A. N.;Hering, B. J.
通讯作者：
Hering, B. J.
影响因子：
3.3
作者：
Lakey, JRT;Warnock, GL;Rajotte, RV
通讯作者：
Rajotte, RV
影响因子：
4.1
作者：
Khosla K;Kangas J;Liu Y;Zhan L;Daly J;Hagedorn M;Bischof J
通讯作者：
Bischof J
影响因子：
2.7
作者：
JUTTE, NHPM;HEYSE, P;ZEILMAKER, GH
通讯作者：
ZEILMAKER, GH