A new method to value efficiency of enzyme blends for pancreatic tissue digestion.
Islet transplantation, since the 1990s, has been an example of human cell therapy. Nevertheless, the islet isolation procedure is not completely standardized; in fact, >50% of islet procedures do not eventuate in transplantation due both to the variability of a donor's pancreas and to the unpredictable efficiency of an enzymatic blend. The enzymes used in pancreas isolation to digest several substrates are extracted from Clostridium histolyticum. In particular, they have strong collagenolytic activity compared with vertebrate collagenases. However, several impediments persist in human islet isolation success, probably owing to the variable composition and concentration of collagenases employed during the digestion phase. For islet isolation processes, neutral proteases play important roles. However, they should be considered to be double-edged swords, contributing to tissue dissociation but, sometimes, decreasing islet yield through fragmentation, breakdown, and inactivation. Protease activities cannot be preciously adjusted in a narrow range, there is no approach to determine the optimal dosage and composition of enzymes for extraction of human islets from the pancreas. At this time, available data on commercial enzymatic activity are not sufficient to predict their efficiency for pancreas digestion; consequently, it is difficult to select enzyme batches. For these reasons, we sought to generate an innovative evaluation assay to select enzymes useful for isolation procedures of pancreatic islets.
Salamone
M
M
IAMC-CNR, U.O. Capo Granitola, Mazara del Vallo, Trapani, Italy. monica.salamone@irma.pa.cnr.it
Seidita
G
G
Cuttitta
A
A
Rigogliuso
S
S
Mazzola
S
S
Bertuzzi
F
F
Ghersi
G
G
D002460:Cell Line; D002469:Cell Separation / Q000379:methods; D003094:Collagen / Q000302:isolation & purification; D017364:Collagenases / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology / Q000502:physiology; D018093:Gelatinases / Q000378:metabolism; D005782:Gels; D006801:Humans; D007515:Islets of Langerhans / Q000166:cytology / Q000201:enzymology*; D016381:Islets of Langerhans Transplantation / Q000379:methods* / Q000502:physiology; D010179:Pancreas / Q000166:cytology / Q000201:enzymology* / Q000502:physiology; D010447:Peptide Hydrolases / Q000378:metabolism; D013820:Thermolysin / Q000378:metabolism
D005782:Gels; D003094:Collagen; D010447:Peptide Hydrolases; D017364:Collagenases; D018093:Gelatinases; D013820:Thermolysin