Rapid purification of islets using magnetic microspheres coated with anti-acinar cell monoclonal antibodies.
A simple, rapid method of islet purification is important in large-scale human islet isolation. We have previously identified monoclonal antibodies specific for acinar cells, but not islets, and described an immunologic method of purification by selective lysis of the acinar cells. An attractive alternative to lysis of the acinar cell is depletion by a magnetic immunomicrosphere technique. We report in this study a rapid, reproducible method of rat islet purification utilizing magnetic microspheres coated with acinar-cell-specific monoclonal antibodies. Pancreatic digestion with collagenase followed by depletion of acinar cells with the magnetic immunomicrospheres (MIMS) yields large numbers of intact islets. We compared the islets thus obtained with hand-picked (HP) islets (control) for yield, purity, in vitro insulin secretory capacities, and in vivo functional viability. The islet yield with the MIMS method (n = 35) was 72.7% that obtained with the HP method (n = 6) (378 +/- 8 vs. 519 +/- 31 islets per pancreas). The purity of the MIMS-isolated islets was 84 +/- 1.9%, ranging from 75-95%. Static glucose stimulation showed excellent function (2-3-fold increase of insulin release over basal levels) with no statistical difference in insulin secretion between MIMS and HP islets. Under microscopic examination, both groups revealed a well-preserved structure with healthy endocrine cells. When 1321 +/- 59 MIMS islets were transplanted into streptozotocin-induced diabetic rats (n = 10), normoglycemia (less than 200 mg/dl) was restored in all recipients following transplantation, and 100% of them remained normoglycemic on day 120 postgrafting. In summary, a rapid, consistent, and simple method of isolating viable, purified rat islets is described. The broad interspecies crossreactivity of the McAb suggests that this technique may be generally useful for islet purification in large mammalia, including man.
Department of Surgery, UCLA School of Medicine.
T
T
Fujioka
P I
PI
Terasaki
R
R
Heintz
N
N
Merideth
R P
RP
Lanza
T L
TL
Zheng
P
P
Soon-Shiong
D000818:Animals; D000911:Antibodies, Monoclonal; D002469:Cell Separation / Q000379:methods; D007515:Islets of Langerhans / Q000166:cytology; D008280:Magnetics; D008863:Microspheres; D010179:Pancreas / Q000166:cytology / Q000276:immunology; D051381:Rats; D011917:Rats, Inbred Lew
D000911:Antibodies, Monoclonal