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Histopaque provides optimal mouse islet purification kinetics: comparison study with Ficoll, iodixanol and dextran.

Authors :
McCall MD
Maciver AH
Pawlick R
Edgar R
Shapiro AM
Source :
Islets [Islets] 2011 Jul-Aug; Vol. 3 (4), pp. 144-9. Date of Electronic Publication: 2011 Jul 01.
Publication Year :
2011

Abstract

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.

Details

Language :
English
ISSN :
1938-2022
Volume :
3
Issue :
4
Database :
MEDLINE
Journal :
Islets
Publication Type :
Academic Journal
Accession number :
21606673
Full Text :
https://doi.org/10.4161/isl.3.4.15729