Optimization of neutral protease to collagenase activity ratio for islet of langerhans isolation

Islet autotransplantation (IAT) is used to preserve as much insulin-secretory capacity as possible in patients undergoing total pancreatectomy for painful chronic pancreatitis. The enzyme used to dissociate the pancreas is a critical determinant of islet yield, which is correlated with posttransplant function. Here, we present our experience with IAT procedures to compare islet product data using the new enzyme SERVA/Nordmark (SN group; n = 46) with the standard enzyme Liberase-HI (LH group; n = 40). Total islet yields (mean ± standard deviation; 216 417 ± 79 278 islet equivalent [IEQ] in the LH group; 227 958 ± 58 544 IEQ in the SN group; p = 0.67) were similar. However, the percentage of embedded islets is higher in the SN group compared to the LH group. Significant differences were found in pancreas digestion time, dilution time, and digested pancreas weight between the two groups. Multivariate linear regression analysis showed the two groups differed in portal venous pressure changes. The incidence of graft function and insulin independence was not different between the two groups. The SN and LH enzymes are associated with similar outcomes for IAT. Further optimization of the collagenase/neutral protease ratio is necessary to reduce the number of embedded islets obtained when using the SN enzyme.

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“…(27,28), donor BMI (23), islet isolation technique (28) and type of enzyme used for pancreas dissociation (13,26 …”

Section: We Have Successfully Utilized the New Enzyme Sn For Autologomentioning

confidence: 99%

Human Islet Isolation for Autologous Transplantation: Comparison of Yield and Function Using SERVA/Nordmark Versus Roche Enzymes

Anazawa

Balamurugan

Bellin

et al. 2009

American Journal of Transplantation

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“…Proteases have been shown to decrease the islet yield in humans through excessive fragmentation and disintegration of pancreas (Bucher et al, 2004). It is therefore possible that this shortcoming accounted for the several islets without preserved morphology observed with protocols incorporating accutase.…”

Section: Insulin Mrna Quantities In Isolated Isletsmentioning

confidence: 99%

Assessment of six different collagenase-based methods to isolate feline pancreatic islets

Zini

Franchini

Guscetti

et al. 2009

Research in Veterinary Science

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“…23 On the basis Pancreatic tissue derived protease activity was assessed in another series of 39 human islet isolations with the standard digestion method using Liberase HI (Roche Applied Science, Indianapolis, IN). Figure 1 shows the changes in protease activity before and after intraductal perfusion of the pancreas using Liberase HI.…”

Section: Resultsmentioning

confidence: 99%

Experience of islet isolation without neutral protease supplementation

Kin¹,

O’Gorman²,

Senior³

et al. 2010

Islets

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Optimization of neutral protease to collagenase activity ratio for islet of langerhans isolation

Cited by 30 publications

References 4 publications

Human Islet Isolation for Autologous Transplantation: Comparison of Yield and Function Using SERVA/Nordmark Versus Roche Enzymes

Human Islet Isolation for Autologous Transplantation: Comparison of Yield and Function Using SERVA/Nordmark Versus Roche Enzymes

Assessment of six different collagenase-based methods to isolate feline pancreatic islets

Experience of islet isolation without neutral protease supplementation

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