Sa. White et al., A preliminary study of the activation of endogenous pancreatic exocrine enzymes during automated porcine islet isolation, CELL TRANSP, 8(3), 1999, pp. 265-276
The activation of endogenous pancreatic enzymes during automated pancreas d
igestion may be detrimental to islet isolation. In this report we assessed
the activation of trypsin, chymotrypsin, elastase, carboxypeptidases A and
B, phospholipase A(2), and lipase using a porcine model. Four islet isolati
ons were examined. Duplicate aliquots were taken from the automated circuit
at 5-min time intervals up to the completion of pancreas digestion (approx
60 min). One aliquot was activated in vitro with exogenous trypsin in orde
r to convert the enzymes into their active non-"proform," with the exceptio
n of trypsinogen, which was activated with exogenous enterokinase. This was
done to assess the percentage activation of each individual enzyme (total
potentially activatable enzyme release). The extent of activation between i
solations was extremely variable. During the closed (recirculating) circuit
phase of pancreas digestion there were both gradual and rapid increases in
the levels of enzymes released. Peak activity of enzyme activation varied
from 13 to 30 min; similarly, total potentially activatable peaks occurred
between 13 and 38 min. Lipase and carboxypeptidase B showed greater than 70
% activation, chymotrypsin, carboxypeptidase A, and phospholipase A, betwee
n 50% and 70% activation, and trypsin and elastase less than 20%. There wer
e up to 30-fold differences between the four islet preparations. In summary
, it is unlikely that poor islet yields are soley explained by variations b
etween collagenases; the variable activation of endogenous pancreatic exocr
ine enzymes is also likely to be influential to porcine islet yields.