Assessment of human islet isolation with four different collagenases.
BACKGROUND
The isolation of islets from the human pancreas critically depends on the efficiency of the digestive enzymes. Liberase HI had been used as a standard preparation until the issues concerning bovine spongiform encephalopathy. Thus, we must now use other collagenases for clinical islet transplantation, four of which we have evaluated herein.


METHODS
The digestion of each of 17 pancreata from brain-dead donors was performed using the following collagenases: Liberase HI (HI; Roche, n = 9); Liberase MTF C/T (MTF; Roche, n = 4); Collagenase NB1 Premium Grade (NB1; Serva, n = 7); or Clzyme Collagenase HA (CI, VitaCyte, n = 4). Islet isolations were based on the Edmonton protocol for HI, whereas our modified islet isolation method was used for the three new enzymes (MTF, NB1, and CI).


RESULTS
There were no significant differences in donor age, body mass index, pancreas size, and cold ischemic time among the four groups. The phase I time in the NB1 group was significantly shorter than in the CI group (P = .0014). The prepurification IEQ/g in the HI group was significantly lower than the others (P = .0003 vs MTF, .0007 vs NB1, and .0009 vs CI, respectively). The postpurification IEQ/g in the MTF group was significantly higher than in the HI group (P = .006). The viability in the NB1 group was significantly greater than the HI group (P = .003).


CONCLUSIONS
Three new enzymes (MTF, NB1, and CI) may enable us to obtain higher islet yields than with HI.
Shimoda
M
M
Division of Cardiology, Department of Internal Medicine, Baylor University Medical Center, Baylor Heart and Vascular Institute, Dallas, Texas, USA.
Noguchi
H
H
Naziruddin
B
B
Fujita
Y
Y
Chujo
D
D
Takita
M
M
Peng
H
H
Tamura
Y
Y
Olsen
G S
GS
Sugimoto
K
K
Itoh
T
T
Onaca
N
N
Levy
M F
MF
Grayburn
P A
PA
Matsumoto
S
S
D000328:Adult; D000818:Animals; D015992:Body Mass Index; D001926:Brain Death; D002417:Cattle; D002469:Cell Separation / Q000379:methods*; D017364:Collagenases / Q000378:metabolism*; D016643:Encephalopathy, Bovine Spongiform / Q000473:pathology; D006801:Humans; D007515:Islets of Langerhans / Q000166:cytology* / Q000473:pathology; D008875:Middle Aged; D009929:Organ Size; D010179:Pancreas / Q000033:anatomy & histology / Q000473:pathology; D013820:Thermolysin / Q000378:metabolism; D014019:Tissue Donors
D017364:Collagenases; C106635:Liberase; D013820:Thermolysin