PRISM: A Novel Human Islet Isolation Technique

被引:2
|
作者
Doppenberg, Jason B. [1 ,2 ]
Engelse, Marten A. [1 ]
de Koning, Eelco J. P. [1 ]
机构
[1] Leiden Univ, Dept Internal Med, Med Ctr, Leiden, Netherlands
[2] Leiden Univ, Transplantat Ctr, Med Ctr, Leiden, Netherlands
关键词
PANCREATIC DIGEST; DENSITY GRADIENT; PORCINE ISLETS; SINGLE-DONOR; CELL SALVAGE; FAT REMOVAL; UW SOLUTION; IN-SITU; PURIFICATION; TRANSPLANTATION;
D O I
10.1097/TP.0000000000003897
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Background. Successful pancreatic islet isolations are a key requirement for islet transplantation in selected patients with type 1 diabetes. However, islet isolation is a technically complex, time-consuming, and manual process. Optimization and simplification of the islet isolation procedure could increase islet yield and quality, require fewer operators, and thus reduce cost. Methods. We developed a new, closed system of tissue collection, washing, buffer change, and islet purification termed PancReatic Islet Separation Method (PRISM). In the developmental phase, pump and centrifuge speed was tested using microspheres with a similar size, shape, and density as digested pancreatic tissue. After optimization, PRISM was used to isolate islets from 10 human pancreases. Results. Islet equivalents viability (fluorescein diacetate/propidium iodide), morphology, and dynamic glucose-stimulated insulin secretion were evaluated. PRISM could be performed by 1 operator in 1 flow cabinet. A similar islet yield was obtained using PRISM compared to the traditional islet isolation method (431 234 +/- 292 833 versus 285 276 +/- 197 392 islet equivalents, P = 0.105). PRISM islets had similar morphology and functionality. Conclusions. PRISM is a novel islet isolation technique that can significantly improve islet isolation efficiency using fewer operators.
引用
收藏
页码:1271 / 1278
页数:8
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