Islet filtration: a simple and rapid new purification procedure that avoids ficoll and improves islet mass and function.

PRO Salvalaggio, S Deng, CE Ariyan, I Millet… - …, 2002 - journals.lww.com
PRO Salvalaggio, S Deng, CE Ariyan, I Millet, WS Zawalich, GP Basadonna, DM Rothstein
Transplantation, 2002journals.lww.com
Islet isolation is a time-consuming process. Islet yields vary, and previous in vitro studies
suggest that Ficoll may be an islet toxin. Here, we describe an alternative, Ficoll-free method
to purify murine islets by filtration through a cell strainer. Collagenase digestion of pancreata
was carried out using standard procedures. The pancreatic digest was divided into aliquots
and purified either by Ficoll or by filtration. Following filtration, islets were intact and
separated from nondigested tissue. Purity was similar to that achieved using Ficoll …
Abstract
Islet isolation is a time-consuming process. Islet yields vary, and previous in vitro studies suggest that Ficoll may be an islet toxin. Here, we describe an alternative, Ficoll-free method to purify murine islets by filtration through a cell strainer. Collagenase digestion of pancreata was carried out using standard procedures. The pancreatic digest was divided into aliquots and purified either by Ficoll or by filtration. Following filtration, islets were intact and separated from nondigested tissue. Purity was similar to that achieved using Ficoll. However, purification by filtration was faster, increased islet yield, and resulted in higher insulin secretion in vitro. Moreover, when syngeneic diabetic hosts were transplanted with a marginal islet mass, islets purified by filtration restored normoglycemia significantly faster than those isolated by Ficoll. This suggests that Ficoll exposure negatively impacts islet function. In conclusion, islet filtration is a simple and rapid procedure for purification of islets that demonstrate improved functional mass.
Lippincott Williams & Wilkins