The development of new density gradient media for purifying human islets and islet-quality assessments

被引:102
作者
Huang, GC
Zhao, M
Jones, P
Persaud, S
Ramracheya, R
Lobner, K
Christie, MR
Banga, JP
Peakman, M
Sirinivsan, P
Rela, M
Heaton, N
Amiel, S
机构
[1] Kings Coll London, GKT Med Sch, Dept Diabet Endocrinol & Internal Med, London SE5 9PJ, England
[2] Kings Coll London, GKT Sch Biomed Sci, Ctr Reprod Endocrinol & Diabet, London SE5 9PJ, England
[3] Kings Coll London, GKT Sch Med & Dent, Rayne Inst, Dept Immunol, London SE5 9PJ, England
[4] Kings Coll London, GKT Med Sch, Inst Liver Studies, London SE5 9PJ, England
关键词
D O I
10.1097/01.TP.0000100401.62912.B2
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 [免疫学];
摘要
Successful islet transplantation is dependent on the quality and quantity of islets infused. Islets are purified on density gradients, but procedures currently used have limited capacity for pancreatic digests, islet yield, and viability. We aimed to improve islet purification with a modified gradient medium. Biocoll was diluted in University of Wisconsin solution to create linear density gradients of 1.065 to 1.095 g/mL. Properties of islets purified from 22 human pancreas digests with modified medium were compared with 15 preparations using standard medium. The modification increased the capacity of gradients for pancreatic digests from 20 to 60 mL, islet yield increased from 218,000 to 435,318 per isolation, and viability increased from 65.4% to 92.1%. Islet fractions contained greater than 95% of recovered insulin. Islets showed good physiologic responses to secretagogues and restored normoglycemia in streptozotocin-induced diabetic severe combined immunodeficiency disease mice. The new medium enhances yield, purity, and viability of human islet preparations for clinical islet transplantation.
引用
收藏
页码:143 / 145
页数:3
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