A new enzyme mixture to increase the yield and transplant rate of autologous and allogeneic human islet products

A. N. Balamurugan, Gopalakrishnan Loganathan, Melena D. Bellin, Joshua J. Wilhelm, James Harmon, Takayuki Anazawa, Sajjad M. Soltani, David M. Radosevich, Takeshi Yuasa, Mukesh Tiwari, Klearchos K Papas, Robert McCarthy, David E R Sutherland, Bernhard J. Hering

Research output: Contribution to journalArticle

61 Citations (Scopus)

Abstract

Background. The optimal enzyme blend that maximizes human islet yield for transplantation remains to be determined. In this study, we evaluated eight different enzyme combinations (ECs) in an attempt to improve islet yield. The ECs consisted of purified, intact or truncated class 1 (C1) and class 2 (C2) collagenases from Clostridium histolyticum (Ch), and neutral protease (NP) from Bacillus thermoproteolyticus rokko (thermolysin) or Ch (ChNP). Methods. We report the results of 249 human islet isolations, including 99 deceased donors (research n=57, clinical n=42) and 150 chronic pancreatitis pancreases. We prepared a new enzyme mixture (NEM) composed of intact C1 and C2 collagenases and ChNP in place of thermolysin. The NEM was first tested in split pancreas (n=5) experiments and then used for islet autologous (n=21) and allogeneic transplantation (n=10). Islet isolation outcomes from eight different ECs were statistically compared using multivariate analysis. Results. The NEM consistently achieved higher islet yields from pancreatitis (P<0.003) and deceased donor pancreases (P<0.001) than other standard ECs. Using the NEM, islet products met release criteria for transplantation from 8 of 10 consecutive pancreases, averaging 6510±2150 islet equivalent number/gram (IEQ/g) pancreas and 694,681±147,356 total IEQ/transplantation. In autologous isolation, the NEM yielded more than 200,000 IEQ from 19 of 21 pancreases (averaging 422,893±181,329 total IEQ and 5979±1469 IEQ/kg recipient body weight) regardless of the severity of fibrosis. Conclusions. A NEM composed of ChNP with CIzyme high intact C1 collagenase recovers higher islet yield from deceased and pancreatitis pancreases while retaining islet quality and function.

Original languageEnglish (US)
Pages (from-to)693-702
Number of pages10
JournalTransplantation
Volume93
Issue number7
DOIs
StatePublished - Apr 15 2012
Externally publishedYes

Fingerprint

Autografts
Pancreas
Enzymes
Thermolysin
Pancreatitis
Clostridium histolyticum
Microbial Collagenase
Transplantation
Matrix Metalloproteinase 8
Islets of Langerhans Transplantation
Homologous Transplantation
Chronic Pancreatitis
Bacillus
Fibrosis
Peptide Hydrolases
Multivariate Analysis
Body Weight

Keywords

  • Autotransplantation
  • Collagenase
  • Human islets
  • Islet isolation
  • Neutral protease
  • Pancreatitis

ASJC Scopus subject areas

  • Transplantation

Cite this

Balamurugan, A. N., Loganathan, G., Bellin, M. D., Wilhelm, J. J., Harmon, J., Anazawa, T., ... Hering, B. J. (2012). A new enzyme mixture to increase the yield and transplant rate of autologous and allogeneic human islet products. Transplantation, 93(7), 693-702. https://doi.org/10.1097/TP.0b013e318247281b

A new enzyme mixture to increase the yield and transplant rate of autologous and allogeneic human islet products. / Balamurugan, A. N.; Loganathan, Gopalakrishnan; Bellin, Melena D.; Wilhelm, Joshua J.; Harmon, James; Anazawa, Takayuki; Soltani, Sajjad M.; Radosevich, David M.; Yuasa, Takeshi; Tiwari, Mukesh; Papas, Klearchos K; McCarthy, Robert; Sutherland, David E R; Hering, Bernhard J.

In: Transplantation, Vol. 93, No. 7, 15.04.2012, p. 693-702.

Research output: Contribution to journalArticle

Balamurugan, AN, Loganathan, G, Bellin, MD, Wilhelm, JJ, Harmon, J, Anazawa, T, Soltani, SM, Radosevich, DM, Yuasa, T, Tiwari, M, Papas, KK, McCarthy, R, Sutherland, DER & Hering, BJ 2012, 'A new enzyme mixture to increase the yield and transplant rate of autologous and allogeneic human islet products', Transplantation, vol. 93, no. 7, pp. 693-702. https://doi.org/10.1097/TP.0b013e318247281b
Balamurugan, A. N. ; Loganathan, Gopalakrishnan ; Bellin, Melena D. ; Wilhelm, Joshua J. ; Harmon, James ; Anazawa, Takayuki ; Soltani, Sajjad M. ; Radosevich, David M. ; Yuasa, Takeshi ; Tiwari, Mukesh ; Papas, Klearchos K ; McCarthy, Robert ; Sutherland, David E R ; Hering, Bernhard J. / A new enzyme mixture to increase the yield and transplant rate of autologous and allogeneic human islet products. In: Transplantation. 2012 ; Vol. 93, No. 7. pp. 693-702.
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AU - Balamurugan, A. N.

AU - Loganathan, Gopalakrishnan

AU - Bellin, Melena D.

AU - Wilhelm, Joshua J.

AU - Harmon, James

AU - Anazawa, Takayuki

AU - Soltani, Sajjad M.

AU - Radosevich, David M.

AU - Yuasa, Takeshi

AU - Tiwari, Mukesh

AU - Papas, Klearchos K

AU - McCarthy, Robert

AU - Sutherland, David E R

AU - Hering, Bernhard J.

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AB - Background. The optimal enzyme blend that maximizes human islet yield for transplantation remains to be determined. In this study, we evaluated eight different enzyme combinations (ECs) in an attempt to improve islet yield. The ECs consisted of purified, intact or truncated class 1 (C1) and class 2 (C2) collagenases from Clostridium histolyticum (Ch), and neutral protease (NP) from Bacillus thermoproteolyticus rokko (thermolysin) or Ch (ChNP). Methods. We report the results of 249 human islet isolations, including 99 deceased donors (research n=57, clinical n=42) and 150 chronic pancreatitis pancreases. We prepared a new enzyme mixture (NEM) composed of intact C1 and C2 collagenases and ChNP in place of thermolysin. The NEM was first tested in split pancreas (n=5) experiments and then used for islet autologous (n=21) and allogeneic transplantation (n=10). Islet isolation outcomes from eight different ECs were statistically compared using multivariate analysis. Results. The NEM consistently achieved higher islet yields from pancreatitis (P<0.003) and deceased donor pancreases (P<0.001) than other standard ECs. Using the NEM, islet products met release criteria for transplantation from 8 of 10 consecutive pancreases, averaging 6510±2150 islet equivalent number/gram (IEQ/g) pancreas and 694,681±147,356 total IEQ/transplantation. In autologous isolation, the NEM yielded more than 200,000 IEQ from 19 of 21 pancreases (averaging 422,893±181,329 total IEQ and 5979±1469 IEQ/kg recipient body weight) regardless of the severity of fibrosis. Conclusions. A NEM composed of ChNP with CIzyme high intact C1 collagenase recovers higher islet yield from deceased and pancreatitis pancreases while retaining islet quality and function.

KW - Autotransplantation

KW - Collagenase

KW - Human islets

KW - Islet isolation

KW - Neutral protease

KW - Pancreatitis

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