We investigated the effect of the activation of complement in human se
rum on isolated adult porcine islets using an in vitro model of pig-to
-human islet transplantation. Pancreata were obtained from slaughterho
use pigs (6-8 mo old). Islets were prepared by intraductal collagenase
digestion followed by purification on Ficoll gradients. The purified
islets were incubated with xenogeneic human serum with or without heat
inactivation for 45 min. As control, islets were incubated with autol
ogous porcine serum. After the incubation, the islets' responsiveness
to an acute glucose stimulus (5.5 mM, static incubation) was evaluated
by measurement of the insulin content of the medium. Islets exposed t
o human serum showed significantly lower insulin secretory response th
an the control (1.76 +/- 1.17 mu U/islet/120 min, without heat inactiv
ation; 1.74 +/- 1.36 mu U/islet/120 min, with heat inactivation; 3.39
+/- 0.92 mu U/ islet/120 min, control). No difference in insulin secre
tory response, however, was observed between islets exposed to human s
erum with heat inactivation and without. Furthermore, we evaluated the
cytotoxic activity of human serum on porcine islets by a complement-d
ependent cytotoxicity using the MTT colorimetric assay, and found that
the human serum had no complement-dependent cytotoxic activity agains
t the islets. We concluded that the insulin secretion dysfunction of p
orcine islets exposed to human serum was not due to the activation of
complement and there was no evidence of hyperacute rejection mediated
by complement activation in the in vitro model of pig-to-human islet t
ransplantation.