scholarly journals Pancreatic Islet Purification from Large Mammals and Humans Using a COBE 2991 Cell Processor versus Large Plastic Bottles

2020 ◽  
Vol 10 (1) ◽  
pp. 10
Author(s):  
Hirofumi Noguchi
Keyword(s):  
Shear Force ◽  
Mechanical Damage ◽  
Islet Isolation ◽  
Cell Processor ◽  
Large Mammals ◽  
Purification Step ◽  
Purification Method ◽  
Continuous Density ◽  
Porcine Islet ◽  
Intraportal Infusion

The islet purification step in clinical islet isolation is important for minimizing the risks associated with intraportal infusion. Continuous density gradient with a COBE 2991 cell processor is commonly used for clinical islet purification. However, the high shear force involved in the purification method using the COBE 2991 cell processor causes mechanical damage to the islets. We and other groups have shown human/porcine islet purification using large cylindrical plastic bottles. Shear stress can be minimized or eliminated using large cylindrical plastic bottles because the bottles do not have a narrow segment and no centrifugation is required during tissue loading and the collection processes of islet purification. This review describes current advances in islet purification from large mammals and humans using a COBE 2991 cell processor versus large cylindrical plastic bottles.

scholarly journals Comparison of Tissue Loading Before and After the Creation of a Continuous Density Gradient in Porcine Islet Purification

Cell Medicine ◽  
2018 ◽  
Vol 10 ◽  
pp. 215517901878134 ◽  
Author(s):  
Chika Miyagi-Shiohira ◽  
Yoshiki Nakashima ◽  
Nana Ebi ◽  
Eri Hamada ◽  
Yoshihito Tamaki ◽  
...  
Keyword(s):  
Density Gradient ◽  
Stimulation Index ◽  
High Yield ◽  
Pancreatic Islet Transplantation ◽  
Post Transplantation ◽  
Porcine Pancreas ◽  
Purification Method ◽  
Continuous Density ◽  
Porcine Islet ◽  
The Impact

The purification step is one of the most important and difficult procedures in islet isolation for pancreatic islet transplantation. We previously reported that a purification method using large plastic bottles effectively achieved a high yield of islets from the porcine pancreas. In this study, we evaluated the impact of the timing of tissue loading on porcine islet purification using large plastic bottles. One method involved loading digested tissue after creating a continuous density gradient (tissue after gradient [TAG]). The other method involved loading digested tissue before creating a continuous density gradient (tissue before gradient [TBG]). There were no significant differences between TAG and TBG in terms of the islet yield, rates of viability and purity, score, and in the stimulation index after purification. Furthermore, there were no marked differences in the attainability or suitability of post-transplantation normoglycemia. Our study shows the equivalency of these two methods of islet purification.

scholarly journals Comparison of Purification Solutions with Different Osmolality for Porcine Islet Purification

Cell Medicine ◽  
2017 ◽  
Vol 9 (1-2) ◽  
pp. 53-59 ◽  
Author(s):  
Chika Miyagi-Shiohira ◽  
Naoya Kobayashi ◽  
Issei Saitoh ◽  
Masami Watanabe ◽  
Yasufumi Noguchi ◽  
...  
Keyword(s):  
Salt Solution ◽  
Stimulation Index ◽  
Human Islets ◽  
Human Islet ◽  
Low Density ◽  
Islet Isolation ◽  
Continuous Density ◽  
Yield Rate ◽  
Purification Efficiency ◽  
Porcine Islet

The osmolality of the purification solution is one of the most critical variables in human islet purification during islet isolation. We previously reported the effectiveness of a combined continuous density/osmolality gradient for the supplemental purification of human islets. We herein applied a combined continuous density/osmolality gradient for regular purification. The islets were purified with a continuous density gradient without osmolality preparation [continuous density/normal osmolality (CD/NO)] or continuous density/osmolality solution with osmolality preparation by 10× Hank's balanced salt solution (HBSS) [continuous density/continuous osmolality (CD/CO)]. The osmolality of the low-density solution was 400 mOsm/kg in both groups and that of the high-density solution was 410 mOsm/kg in the CD/NO group and 500 mOsm/kg in the CD/CO group. Unexpectedly, we noted no significant differences between the two solutions in terms of the islet yield, rate of viability and purity, score, stimulation index, or the attainability and suitability of posttransplantation normoglycemia. Despite reports that the endocrine and exocrine tissues of pancreata have distinct osmotic sensitivities and that high-osmolality solutions result in greater purification efficiency, the isolation and transplant outcomes did not markedly differ between the two purification solutions with different osmolalities in this study.

scholarly journals Evaluation of Islet Purification Methods for Making a Continuous Density Gradient and Loading Tissue

Cell Medicine ◽  
2018 ◽  
Vol 10 ◽  
pp. 215517901773309 ◽  
Author(s):  
Nana Ebi ◽  
Chika Miyagi-Shiohira ◽  
Eri Hamada ◽  
Yoshihito Tamaki ◽  
Mariko Masamoto ◽  
...  
Keyword(s):  
Density Gradient ◽  
Stimulation Index ◽  
High Yield ◽  
Islet Isolation ◽  
Pancreatic Islet Transplantation ◽  
Porcine Pancreas ◽  
Purification Method ◽  
Continuous Density ◽  
Yield Rate ◽  
Purification Methods

Islet purification is one of the most important steps of islet isolation for pancreatic islet transplantation. We previously reported that a purification method using large plastic bottles effectively achieved a high yield of islets from porcine pancreas. In this study, we evaluated the methods for making a continuous density gradient and loading tissue. One method involved loading digested tissue on top of a continuous density gradient (top loading). The other method involved mixing digested tissue with low-density solution and then making a continuous gradient (mixed loading). There were no significant differences between the 2 purification methods in terms of the islet yield, rate of viability or purity, score, or in the stimulation index after purification. Furthermore, there were no marked differences in the attainability or suitability of posttransplantation normoglycemia. Our study shows the equivalency of these 2 methods of islet purification.

Prevention of oxidative stress in porcine islet isolation

2010 ◽  
Vol 13 (1) ◽  
pp. 38-47 ◽  
Author(s):  
Philipp Stiegler ◽  
Vanessa Stadlbauer ◽  
Florian Hackl ◽  
Silvia Schaffellner ◽  
Florian Iberer ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Islet Isolation ◽  
Porcine Islet

scholarly journals A Preliminary Study of the Activation of Endogenous Pancreatic Exocrine Enzymes during Automated Porcine Islet Isolation

1999 ◽  
Vol 8 (3) ◽  
pp. 265-276 ◽  
Author(s):  
S. A. White ◽  
H. Djaballah ◽  
D. P. Hughes ◽  
D. L. Roberts ◽  
H. H. Contractor ◽  
...  
Keyword(s):  
Islet Isolation ◽  
Porcine Islet ◽  
Pancreatic Exocrine ◽  
Preliminary Study

scholarly journals A Novel Preservation Solution Containing a JNK Inhibitory Peptide Efficiently Improves Islet Yield for Porcine Islet Isolation

2019 ◽  
Vol 103 (2) ◽  
pp. 344-352 ◽  
Author(s):  
Hirofumi Noguchi ◽  
Chika Miyagi-Shiohira ◽  
Yoshiki Nakashima ◽  
Nana Ebi ◽  
Eri Hamada ◽  
...  
Keyword(s):  
Islet Isolation ◽  
Inhibitory Peptide ◽  
Preservation Solution ◽  
Porcine Islet

Development of a novel digestion chamber for human and porcine islet isolation

2004 ◽  
Vol 36 (4) ◽  
pp. 1135-1138 ◽  
Author(s):  
D.W.R Gray ◽  
N Sudhakaran ◽  
T.T Titus ◽  
P McShane ◽  
P Johnson
Keyword(s):  
Islet Isolation ◽  
Porcine Islet

AMOUNT AND DISTRIBUTION OF COLLAGEN IN THE PANCREAS HAVE NO EFFECT ON PORCINE ISLET ISOLATION OUTCOME

2008 ◽  
Vol 86 (Supplement) ◽  
pp. 570-571
Author(s):  
D E. Hilling ◽  
E Bouwman ◽  
J K.R.A. Rijkelijkhuizen ◽  
H A.M. Töns ◽  
O T. Terpstra
Keyword(s):  
Islet Isolation ◽  
Porcine Islet

The Use of Pancreas Biopsy Scoring Provides Reliable Porcine Islet Yields While Encapsulation Permits the Determination of Microbiological Safety

2005 ◽  
Vol 14 (7) ◽  
pp. 427-439 ◽  
Author(s):  
Lawrence S. Gazda ◽  
Hollie Adkins ◽  
Johannah A. Bailie ◽  
Wendy Byrd ◽  
Lisa Circle ◽  
...  
Keyword(s):  
Diabetic Patients ◽  
Islet Isolation ◽  
Isolation Technique ◽  
Porcine Islet ◽  
Porcine Endogenous Retrovirus ◽  
Pancreas Biopsy ◽  
Microbiological Testing ◽  
Islet Size ◽  
Biopsy Score

For clinical xenogenic islet transplantation to be successful, several requirements must be met. Among them is a sizeable and reliable source of fully functional and microbiologically safe islets. The inherent variability among porcine pancreases, with respect to islet yield, prompted us to develop a Biopsy Score technique to determine the suitability of each pancreas for islet isolation processing. The Biopsy Score consists of an assessment of five variables: warm ischemia time, pancreas color, fat content, islet size, and islet demarcation, each of which is assigned a value of −1 or +1, depending on whether or not the established criteria is met. For determination of islet size and demarcation, fresh biopsies of porcine pancreases are stained with dithizone (DTZ) solution and examined under a dissecting microscope. Based on the scoring of such biopsies in pancreases from 26—56-month-old sows, we report here that the presence of large (>100 μm diameter), well-demarcated islets in the pancreas biopsy is a reliable predictor of isolation success. Encapsulation of the isolated porcine islets within the inner layer of a 1.5% agarose and an outer layer of 5.0% agarose macrobead, containing 500 equivalent islet number (EIN), provides for extended in vitro functional viability (>6 months of insulin production in response to glucose), as well as for comprehensive microbiological testing and at least partial isolation of the xenogeneic islets from the host immune system. All microbiological testing to date has been negative, except for the presence of porcine endogenous retrovirus (PERV). Taken together, we believe that the Biopsy Score enhancement of our islet isolation technique and our agarose-agarose macroencapsulation methodology bring us significantly closer to realizing clinical porcine islet xenotransplantation for the treatment of insulin-dependent diabetic patients.

The Effect of UW Solution and Its Components on the Collagenase Digestion of Human and Porcine Pancreas

1995 ◽  
Vol 4 (6) ◽  
pp. 615-619 ◽  
Author(s):  
Harold H. Contractor ◽  
Paul R. V. Johnson ◽  
David R. Chadwick ◽  
Gavin S. M. Robertson ◽  
Nicholas J. M. London
Keyword(s):  
Cold Storage ◽  
Species Difference ◽  
Human Pancreas ◽  
Islet Isolation ◽  
Porcine Pancreas ◽  
Uw Solution ◽  
Porcine Islet ◽  
University Of Wisconsin ◽  
Collagenase Digestion ◽  
Tissue Dissociation

University of Wisconsin (UW) solution is used extensively as a cold storage solution during the procurement and transport of the pancreas prior to islet isolation. However, it has been observed that UW inhibits the collagenase digestion phase of human but not porcine islet isolation, resulting in poor islet yields and islets of poor viability. The aim of this study was, therefore, to confirm this species difference and to determine which components of UW are responsible for the inhibition in the human. In the initial experiment, blocks of human and porcine pancreas (n = 7) were incubated in test tubes containing collagenase at a concentration of 4 mg/mL at 37°C dissolved in 4 mL of either Hanks' solution or UW. Every 5 min the tubes were manually shaken and the degree of tissue dissociation scored on a scale of + and +++. Our results confirm the inhibition of collagenase digestion in the human but not the pig. Using the same methodology, we then investigated the components of UW that were causing the observed inhibition in the human pancreas (n = 7). This time the collagenase was dissolved in individual or combinations of UW components. Using Hank's as a control, the results were then expressed as a median ratio. The components found to be most inhibitory were magnesium, the Na+/K+ ratio, hydroxyethyl starch (HES), and adenosine. Allopurinol in combination with either lactobionate or glutathione was markedly inhibitory (i.e., median ratio 1.8 and 1.9, respectively). The most inhibitory solution tested was a combination of the three components raffinose, glutathione, and lactobionate (median ratio 2.1). This combination was almost as inhibitory as UW itself (median ratio 2.7). These findings are essential for the development of effective cold-storage solutions for the human pancreas that do not inhibit the subsequent collagenase digestion phase of islet isolation.

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