Quadrupole magnetic sorting of porcine islets of Langerhans.
Islet transplantation is emerging as a treatment option for selected patients with type 1 diabetes. Inconsistent isolation, purification, and recovery of large numbers of high-quality islets remain substantial impediments to progress in the field. Removing islets as soon as they are liberated from the pancreas during digestion and circumventing the need for density gradient purification is likely to result in substantially increased viable islet yields by minimizing exposure to proteolytic enzymes, reactive oxygen intermediates, and mechanical stress associated with centrifugation. This study capitalized on the hypervascularity of islets compared with acinar tissue to explore their preferential enrichment with magnetic beads to enable immediate separation in a magnetic field utilizing a quadrupole magnetic sorting. The results demonstrate that (1) preferential enrichment of porcine islets is achievable, but homogeneous bead distribution within the pancreas is difficult to achieve with current protocols; (2) greater than 70% of islets in the dissociated pancreatic tissue were recovered by quadrupole magnetic sorting, but their purity was low; and (3) infused islets purified by density gradients and subsequently passed through quadrupole magnetic sorting had similar potency as uninfused islets. These results demonstrate proof of concept and define the steps for implementation of this technology in pig and human islet isolation.
Shenkman
Rustin M
RM
Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, Ohio, USA.
Chalmers
Jeffrey J
JJ
Hering
Bernhard J
BJ
Kirchhof
Nicole
N
Papas
Klearchos K
KK
D000818:Animals; D002469:Cell Separation / Q000379:methods*; D005947:Glucose / Q000494:pharmacology; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology* / Q000187:drug effects; D008280:Magnetics / Q000379:methods*; D008863:Microspheres; D013194:Staining and Labeling; D034421:Sus scrofa
D007328:Insulin; D005947:Glucose