Optimization of the magnetic field used for immunomagnetic islet purification.

Abstract

Purification of islets based on the physical differences in density between exocrine and islet tissue reduces islet yields and remains one of the factors limiting islet transplantation. Immunomagnetic cell separation methods provide an attractive, highly specific alternative capable of rapid, gentle, high volume cell separation, but they require modification to be applied effectively to separation of the much larger tissue fragments involved in islet purification. In this study, mAb to rat exocrine tissue were coupled to 4.5-microns magnetic beads (M450 Dynabeads), before incubation with standard aliquots of rat pancreatic digest. The effect on immunomagnetic islet purification of modifications in the magnetic field and the method of digest release into the field were investigated. The results showed that using vibration to maintain the immunomagnetically labeled digest in suspension in tissue culture medium whose density had been increased by the addition of BSA, significantly improved the purification process. When the digest suspension was slowly released and allowed to drift under gravity through a magnetic field applied across a narrow tube, the use of a quadripole of permanent magnets improved results compared with bipolar or unipolar magnetic fields. By modifying immunomagnetic cell separation techniques in this way, a median islet yield of 77% could be reliably achieved while removing 88% of the contaminating exocrine tissue. The use of such methods in human islet purification would significantly increase the yield of islets from each donor pancreas and increase the success rate of transplantation from single donors.