Histopaque provides optimal mouse islet purification kinetics Comparison study with Ficoll, iodixanol and dextran

被引:20
作者
McCall, Michael D. [1 ]
Maciver, Allison H. [1 ]
Pawlick, Rena [1 ]
Edgar, Ryan [1 ]
Shapiro, A. M. James [1 ,2 ]
机构
[1] Univ Alberta, Dept Surg, Edmonton, AB, Canada
[2] Univ Alberta, Clin Islet Transplant Program, Edmonton, AB, Canada
关键词
islets of langerhans; gradient; histopaque; ficoll; islet isolation; DISSEMINATED INTRAVASCULAR COAGULATION; RAT PANCREATIC-ISLETS; BOVINE SERUM-ALBUMIN; DENSITY GRADIENT; PORTAL-HYPERTENSION; TRANSPLANTATION; LANGERHANS; SEPARATION; DANGER;
D O I
10.4161/isl.3.4.15729
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Islet transplantation has become a very promising treatment for type 1 diabetes. To facilitate further clinical improvements in this exciting field, rodent islets are used to evaluate new strategies and modifications. One method to purify islets is on a density gradient, although the optimal gradient component can be debated. N = 6 separate mouse islet isolations were used and the resulting islets were separated and purified on either a Ficoll, Histopaque, dextran or iodixanol gradient. Islets were assessed for recovery, viability, purity and in vitro functionality. Aliquots were transplanted into diabetic mice to assess in vivo functionality and survival. There was no difference in the number of islets recovered across groups nor in the size of recovered islets. Use of a Ficoll or Histopaque gradient led to the most pure and viable islets in comparison to dextran and iodixanol. Functionally, islets isolated on a Ficoll gradient had the highest glucose-stimulated insulin release in vitro while performing equally to Histopaque and dextran gradients in vivo. Using a Ficoll gradient, however, comes at a higher monetary cost. We recommend using a Histopaque gradient, which led to the isolation of viable and functional islets with a reduced cost as compared to a Ficoll gradient.
引用
收藏
页码:144 / 149
页数:6
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