Surgical protocol involving the infusion of paramagnetic microparticles for preferential incorporation within porcine islets

M. D. Rizzari, T. M. Suszynski, L. S. Kidder, S. A. Stein, T. D. O'Brien, V. S K Sajja, W. E. Scott, V. A. Kirchner, B. P. Weegman, E. S. Avgoustiniatos, P. W. Todd, D. J. Kennedy, B. E. Hammer, D. E R Sutherland, B. J. Hering, Klearchos K Papas

Research output: Contribution to journalArticle

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Abstract

Introduction Despite significant advances, widespread applicability of islet cell transplantation remains elusive. Refinement of current islet isolation protocols may improve transplant outcomes. Islet purification by magnetic separation has shown early promise. However, surgical protocols must be optimized to maximize the incorporation of paramagnetic microparticles (MP) within a greater number of islets. This study explores the impact of MP concentration and infusion method on optimizing MP incorporation within islets. Methods Five porcine pancreata were procured from donors after cardiac death. Splenic lobes were isolated and infused with varying concentrations of MP (8, 16, and 32 × 10 8 MP/L of cold preservation solution) and using one of two delivery techniques (hanging bag versus hand-syringe). After procurement and infusion, pancreata were stored at 0°C to 4°C during transportation (less than 1 hour), fixed in 10% buffered formalin, and examined by standard magnetic resonance imaging (MRI) and histopathology. Results T2 (*)-weighted MRI showed homogeneous distribution of MP in all experimental splenic lobes. In addition, histologic analysis confirmed that MP were primarily located within the microvasculature of islets (82% to 85%), with few MP present in acinar tissue (15% to 18%), with an average of five to seven MP per islet (within a 5-μm thick section). The highest MP incorporation was achieved at a concentration of 16 × 10 8 MP/L using the hand-syringe technique. Conclusion This preliminary study suggests that optimization of a surgical protocol, MP concentrations, and applied infusion pressures may enable more uniform distribution of MP in the porcine pancreas and better control of MP incorporation within islets. These results may have implications in maximizing the efficacy of islet purification by magnetic separation.

Original languageEnglish (US)
Pages (from-to)4209-4212
Number of pages4
JournalTransplantation Proceedings
Volume42
Issue number10
DOIs
StatePublished - Dec 2010
Externally publishedYes

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Pancreas
Swine
Syringes
Hand
Magnetic Resonance Imaging
Islets of Langerhans Transplantation
Cell Transplantation
Microvessels
Islets of Langerhans
Formaldehyde
Transplants
Pressure

ASJC Scopus subject areas

  • Surgery
  • Transplantation

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Surgical protocol involving the infusion of paramagnetic microparticles for preferential incorporation within porcine islets. / Rizzari, M. D.; Suszynski, T. M.; Kidder, L. S.; Stein, S. A.; O'Brien, T. D.; Sajja, V. S K; Scott, W. E.; Kirchner, V. A.; Weegman, B. P.; Avgoustiniatos, E. S.; Todd, P. W.; Kennedy, D. J.; Hammer, B. E.; Sutherland, D. E R; Hering, B. J.; Papas, Klearchos K.

In: Transplantation Proceedings, Vol. 42, No. 10, 12.2010, p. 4209-4212.

Research output: Contribution to journalArticle

Rizzari, MD, Suszynski, TM, Kidder, LS, Stein, SA, O'Brien, TD, Sajja, VSK, Scott, WE, Kirchner, VA, Weegman, BP, Avgoustiniatos, ES, Todd, PW, Kennedy, DJ, Hammer, BE, Sutherland, DER, Hering, BJ & Papas, KK 2010, 'Surgical protocol involving the infusion of paramagnetic microparticles for preferential incorporation within porcine islets', Transplantation Proceedings, vol. 42, no. 10, pp. 4209-4212. https://doi.org/10.1016/j.transproceed.2010.09.138
Rizzari, M. D. ; Suszynski, T. M. ; Kidder, L. S. ; Stein, S. A. ; O'Brien, T. D. ; Sajja, V. S K ; Scott, W. E. ; Kirchner, V. A. ; Weegman, B. P. ; Avgoustiniatos, E. S. ; Todd, P. W. ; Kennedy, D. J. ; Hammer, B. E. ; Sutherland, D. E R ; Hering, B. J. ; Papas, Klearchos K. / Surgical protocol involving the infusion of paramagnetic microparticles for preferential incorporation within porcine islets. In: Transplantation Proceedings. 2010 ; Vol. 42, No. 10. pp. 4209-4212.
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abstract = "Introduction Despite significant advances, widespread applicability of islet cell transplantation remains elusive. Refinement of current islet isolation protocols may improve transplant outcomes. Islet purification by magnetic separation has shown early promise. However, surgical protocols must be optimized to maximize the incorporation of paramagnetic microparticles (MP) within a greater number of islets. This study explores the impact of MP concentration and infusion method on optimizing MP incorporation within islets. Methods Five porcine pancreata were procured from donors after cardiac death. Splenic lobes were isolated and infused with varying concentrations of MP (8, 16, and 32 × 10 8 MP/L of cold preservation solution) and using one of two delivery techniques (hanging bag versus hand-syringe). After procurement and infusion, pancreata were stored at 0°C to 4°C during transportation (less than 1 hour), fixed in 10{\%} buffered formalin, and examined by standard magnetic resonance imaging (MRI) and histopathology. Results T2 (*)-weighted MRI showed homogeneous distribution of MP in all experimental splenic lobes. In addition, histologic analysis confirmed that MP were primarily located within the microvasculature of islets (82{\%} to 85{\%}), with few MP present in acinar tissue (15{\%} to 18{\%}), with an average of five to seven MP per islet (within a 5-μm thick section). The highest MP incorporation was achieved at a concentration of 16 × 10 8 MP/L using the hand-syringe technique. Conclusion This preliminary study suggests that optimization of a surgical protocol, MP concentrations, and applied infusion pressures may enable more uniform distribution of MP in the porcine pancreas and better control of MP incorporation within islets. These results may have implications in maximizing the efficacy of islet purification by magnetic separation.",
author = "Rizzari, {M. D.} and Suszynski, {T. M.} and Kidder, {L. S.} and Stein, {S. A.} and O'Brien, {T. D.} and Sajja, {V. S K} and Scott, {W. E.} and Kirchner, {V. A.} and Weegman, {B. P.} and Avgoustiniatos, {E. S.} and Todd, {P. W.} and Kennedy, {D. J.} and Hammer, {B. E.} and Sutherland, {D. E R} and Hering, {B. J.} and Papas, {Klearchos K}",
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T1 - Surgical protocol involving the infusion of paramagnetic microparticles for preferential incorporation within porcine islets

AU - Rizzari, M. D.

AU - Suszynski, T. M.

AU - Kidder, L. S.

AU - Stein, S. A.

AU - O'Brien, T. D.

AU - Sajja, V. S K

AU - Scott, W. E.

AU - Kirchner, V. A.

AU - Weegman, B. P.

AU - Avgoustiniatos, E. S.

AU - Todd, P. W.

AU - Kennedy, D. J.

AU - Hammer, B. E.

AU - Sutherland, D. E R

AU - Hering, B. J.

AU - Papas, Klearchos K

PY - 2010/12

Y1 - 2010/12

N2 - Introduction Despite significant advances, widespread applicability of islet cell transplantation remains elusive. Refinement of current islet isolation protocols may improve transplant outcomes. Islet purification by magnetic separation has shown early promise. However, surgical protocols must be optimized to maximize the incorporation of paramagnetic microparticles (MP) within a greater number of islets. This study explores the impact of MP concentration and infusion method on optimizing MP incorporation within islets. Methods Five porcine pancreata were procured from donors after cardiac death. Splenic lobes were isolated and infused with varying concentrations of MP (8, 16, and 32 × 10 8 MP/L of cold preservation solution) and using one of two delivery techniques (hanging bag versus hand-syringe). After procurement and infusion, pancreata were stored at 0°C to 4°C during transportation (less than 1 hour), fixed in 10% buffered formalin, and examined by standard magnetic resonance imaging (MRI) and histopathology. Results T2 (*)-weighted MRI showed homogeneous distribution of MP in all experimental splenic lobes. In addition, histologic analysis confirmed that MP were primarily located within the microvasculature of islets (82% to 85%), with few MP present in acinar tissue (15% to 18%), with an average of five to seven MP per islet (within a 5-μm thick section). The highest MP incorporation was achieved at a concentration of 16 × 10 8 MP/L using the hand-syringe technique. Conclusion This preliminary study suggests that optimization of a surgical protocol, MP concentrations, and applied infusion pressures may enable more uniform distribution of MP in the porcine pancreas and better control of MP incorporation within islets. These results may have implications in maximizing the efficacy of islet purification by magnetic separation.

AB - Introduction Despite significant advances, widespread applicability of islet cell transplantation remains elusive. Refinement of current islet isolation protocols may improve transplant outcomes. Islet purification by magnetic separation has shown early promise. However, surgical protocols must be optimized to maximize the incorporation of paramagnetic microparticles (MP) within a greater number of islets. This study explores the impact of MP concentration and infusion method on optimizing MP incorporation within islets. Methods Five porcine pancreata were procured from donors after cardiac death. Splenic lobes were isolated and infused with varying concentrations of MP (8, 16, and 32 × 10 8 MP/L of cold preservation solution) and using one of two delivery techniques (hanging bag versus hand-syringe). After procurement and infusion, pancreata were stored at 0°C to 4°C during transportation (less than 1 hour), fixed in 10% buffered formalin, and examined by standard magnetic resonance imaging (MRI) and histopathology. Results T2 (*)-weighted MRI showed homogeneous distribution of MP in all experimental splenic lobes. In addition, histologic analysis confirmed that MP were primarily located within the microvasculature of islets (82% to 85%), with few MP present in acinar tissue (15% to 18%), with an average of five to seven MP per islet (within a 5-μm thick section). The highest MP incorporation was achieved at a concentration of 16 × 10 8 MP/L using the hand-syringe technique. Conclusion This preliminary study suggests that optimization of a surgical protocol, MP concentrations, and applied infusion pressures may enable more uniform distribution of MP in the porcine pancreas and better control of MP incorporation within islets. These results may have implications in maximizing the efficacy of islet purification by magnetic separation.

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