Islet purification method using large bottles effectively achieves high islet yield from pig pancreas.
Porcine islets are a promising resource for xenotransplantation. However, low efficacy of islet isolation because of their marked fragility remains a problem. Recently we found that the standard purification method using COBE 2991 cell processor (COBE) with Ficoll density gradient solution  damaged islets mechanically by high shearing force. In this study, we evaluated our new purification method using large plastic bottles for the efficacy of islet purification. Ten porcine pancreata were used. The average warm ischemic time was over 40 min; therefore, these pancreata were considered  to be in a marginal condition. After digestion, the digested tissue was divided into three groups. Each group was purified using either top loading method with bottle (top group) or bottom loading method with bottle (bottom group) or standard COBE method (COBE group). Islet yield per pancreas  weight (IEQ/g) and the rate of postpurification recovery in the top group were significantly higher than the COBE group (top: 8060 ± 1652 IEQ/g, bottom: 4572 ± 614 IE/g, COBE: 3900 ± 734 IE/g. p < 0.02 in top vs. COBE; top percentage of recovery: 99.3 ±  12.3%, bottom: 62.6 ± 8.8%, COBE: 49.5 ± 6.7%, p < 0.02 in top vs. bottom and COBE). The average sizes of purified islets in the top and bottom groups were significantly larger than COBE group (Average diameter top: 156 ± 8 μm, bottom: 147 ± 6 μm,  COBE: 119 ± 6 μm, p < 0.01 in top vs. COBE and in bottom vs. COBE), which indicated that bottle method can reduce shear force during purification. Our new purification using top loading bottle method enabled us to obtain a high yield of porcine islets from marginal pancreata.
Baylor University Medical Center, Dallas, TX, USA.
Masayuki
M
Shimoda
Hirofumi
H
Noguchi
Yasutaka
Y
Fujita
Morihito
M
Takita
Tetsuya
T
Ikemoto
Daisuke
D
Chujo
Bashoo
B
Naziruddin
Marlon F
MF
Levy
Naoya
N
Kobayashi
Paul A
PA
Grayburn
Shinichi
S
Matsumoto
D000818:Animals; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D002470:Cell Survival / Q000187:drug effects; D002499:Centrifugation, Density Gradient; D005362:Ficoll / Q000494:pharmacology; D005942:Gluconates / Q000737:chemistry / Q000494:pharmacology; D006895:Hydroxyethyl Starch Derivatives / Q000737:chemistry / Q000494:pharmacology; D007515:Islets of Langerhans / Q000166:cytology* / Q000187:drug effects; D010179:Pancreas / Q000166:cytology*; D010710:Phosphates / Q000737:chemistry / Q000494:pharmacology; D013552:Swine; D014199:Trehalose / Q000737:chemistry / Q000494:pharmacology
C087059:ET-Kyoto solution; D005942:Gluconates; D006895:Hydroxyethyl Starch Derivatives; D010710:Phosphates; D005362:Ficoll; D014199:Trehalose