Immunomagnetic isolation of islets from the rat pancreas.
Islets were selectively isolated from rat pancreatic digests using magnetic microspheres coated with anti-islet monoclonal antibodies. The isolation process was optimized as a function of bead concentration and time of incubation with the tissue digest. Apparent and normalized islet yields of 92 +/- 6% and 81 +/- 7%, respectively, were obtained by incubating the digests with 10(7) beads/mL for 40 min followed by isolation of the bead-coated islets under a magnetic field. While some fragmentation occurred, the isolation process did not alter islet functionality as demonstrated by an insulin secretion response to glucose stimulation equivalent to that of handpicked controls. The technique is fast, reproducible, and potentially scaleable to larger pancreases as a secondary purification step.
Nandigala
P
P
Department of Chemical Engineering, Iowa State University, Ames 50011, USA.
Chen
T H
TH
Yang
C
C
Hsu
W H
WH
Heath
C
C
D000818:Animals; D000911:Antibodies, Monoclonal; D005947:Glucose / Q000494:pharmacology; D007075:Immunoglobulin M; D018189:Immunomagnetic Separation; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D010179:Pancreas / Q000166:cytology*; D051381:Rats; D017207:Rats, Sprague-Dawley
D000911:Antibodies, Monoclonal; D007075:Immunoglobulin M; D007328:Insulin; D005947:Glucose