Successful human islet isolation and transplantation indicating the importance of class 1 collagenase and collagen degradation activity assay

A. N. Balamurugan, Andrew G. Breite, Takayuki Anazawa, Gopalakrishnan Loganathan, Joshua J. Wilhelm, Klearchos K Papas, Francis E. Dwulet, Robert C. McCarthy, Bernhard J. Hering

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

Background. Purified tissue dissociation enzymes (TDEs) are critical to successful human islet isolation required for clinical transplantation, but little is known about the characteristics of the key enzymes-class I (C1) and class II (C2) collagenase from Clostridium histolyticum-used in these procedures. Here, we show the differences between the C1 collagenase found in purified collagenase products manufactured by three suppliers and the impact of differences in C1 between two suppliers on human islet yield. Methods. Collagenase from Roche, Serva/Nordmark (Uetersen, Germany), and VitaCyte (Indianapolis, IN) were analyzed by analytical high-performance liquid chromatography and collagen degradation activity (CDA), an assay that preferentially detects intact C1 collagenase. Human islet isolations were performed using current standard practices. Results. These studies showed that the highest amount of intact C1 that correlated with a high specific CDA (CDA unit per milligram of protein). The highest specific CDA was found in VitaCyte product followed by the Roche and Serva/Nordmark products. The products of VitaCyte were used successfully for human islet isolation (n=14) with an average final islet yield obtained was 419,100±150,900 islet equivalent number (IEQ) (4147±1759 IEQ/g pancreas). Four of these preparations were used successfully in clinical transplantation procedures. These TDEs gave significantly better results when compared with earlier data where 27 isolations were performed using Serva NB1 collagenase and NB neutral protease where the final islet yield was 217,500±152,400 IEQ (2134±1524 IEQ/g pancreas). Conclusions. These data indicate the importance of intact C1 and the use of the appropriate analytical assays to correlate biochemical characteristics of TDEs to islet quality and yield.

Original languageEnglish (US)
Pages (from-to)954-961
Number of pages8
JournalTransplantation
Volume89
Issue number8
DOIs
StatePublished - Apr 2010
Externally publishedYes

Fingerprint

Islets of Langerhans Transplantation
Collagenases
Collagen
Enzymes
Pancreas
Microbial Collagenase
Transplantation
Germany
Peptide Hydrolases
High Pressure Liquid Chromatography
collagenase 1
Proteins

Keywords

  • Collagenase
  • Collagenase assay
  • Islet isolation
  • Islet transplantation
  • Tissue Dissociation

ASJC Scopus subject areas

  • Transplantation

Cite this

Successful human islet isolation and transplantation indicating the importance of class 1 collagenase and collagen degradation activity assay. / Balamurugan, A. N.; Breite, Andrew G.; Anazawa, Takayuki; Loganathan, Gopalakrishnan; Wilhelm, Joshua J.; Papas, Klearchos K; Dwulet, Francis E.; McCarthy, Robert C.; Hering, Bernhard J.

In: Transplantation, Vol. 89, No. 8, 04.2010, p. 954-961.

Research output: Contribution to journalArticle

Balamurugan, AN, Breite, AG, Anazawa, T, Loganathan, G, Wilhelm, JJ, Papas, KK, Dwulet, FE, McCarthy, RC & Hering, BJ 2010, 'Successful human islet isolation and transplantation indicating the importance of class 1 collagenase and collagen degradation activity assay', Transplantation, vol. 89, no. 8, pp. 954-961. https://doi.org/10.1097/TP.0b013e3181d21e9a
Balamurugan, A. N. ; Breite, Andrew G. ; Anazawa, Takayuki ; Loganathan, Gopalakrishnan ; Wilhelm, Joshua J. ; Papas, Klearchos K ; Dwulet, Francis E. ; McCarthy, Robert C. ; Hering, Bernhard J. / Successful human islet isolation and transplantation indicating the importance of class 1 collagenase and collagen degradation activity assay. In: Transplantation. 2010 ; Vol. 89, No. 8. pp. 954-961.
@article{75f543bb160b4f6e964fee75f1ae1f99,
title = "Successful human islet isolation and transplantation indicating the importance of class 1 collagenase and collagen degradation activity assay",
abstract = "Background. Purified tissue dissociation enzymes (TDEs) are critical to successful human islet isolation required for clinical transplantation, but little is known about the characteristics of the key enzymes-class I (C1) and class II (C2) collagenase from Clostridium histolyticum-used in these procedures. Here, we show the differences between the C1 collagenase found in purified collagenase products manufactured by three suppliers and the impact of differences in C1 between two suppliers on human islet yield. Methods. Collagenase from Roche, Serva/Nordmark (Uetersen, Germany), and VitaCyte (Indianapolis, IN) were analyzed by analytical high-performance liquid chromatography and collagen degradation activity (CDA), an assay that preferentially detects intact C1 collagenase. Human islet isolations were performed using current standard practices. Results. These studies showed that the highest amount of intact C1 that correlated with a high specific CDA (CDA unit per milligram of protein). The highest specific CDA was found in VitaCyte product followed by the Roche and Serva/Nordmark products. The products of VitaCyte were used successfully for human islet isolation (n=14) with an average final islet yield obtained was 419,100±150,900 islet equivalent number (IEQ) (4147±1759 IEQ/g pancreas). Four of these preparations were used successfully in clinical transplantation procedures. These TDEs gave significantly better results when compared with earlier data where 27 isolations were performed using Serva NB1 collagenase and NB neutral protease where the final islet yield was 217,500±152,400 IEQ (2134±1524 IEQ/g pancreas). Conclusions. These data indicate the importance of intact C1 and the use of the appropriate analytical assays to correlate biochemical characteristics of TDEs to islet quality and yield.",
keywords = "Collagenase, Collagenase assay, Islet isolation, Islet transplantation, Tissue Dissociation",
author = "Balamurugan, {A. N.} and Breite, {Andrew G.} and Takayuki Anazawa and Gopalakrishnan Loganathan and Wilhelm, {Joshua J.} and Papas, {Klearchos K} and Dwulet, {Francis E.} and McCarthy, {Robert C.} and Hering, {Bernhard J.}",
year = "2010",
month = "4",
doi = "10.1097/TP.0b013e3181d21e9a",
language = "English (US)",
volume = "89",
pages = "954--961",
journal = "Transplantation",
issn = "0041-1337",
publisher = "Lippincott Williams and Wilkins",
number = "8",

}

TY - JOUR

T1 - Successful human islet isolation and transplantation indicating the importance of class 1 collagenase and collagen degradation activity assay

AU - Balamurugan, A. N.

AU - Breite, Andrew G.

AU - Anazawa, Takayuki

AU - Loganathan, Gopalakrishnan

AU - Wilhelm, Joshua J.

AU - Papas, Klearchos K

AU - Dwulet, Francis E.

AU - McCarthy, Robert C.

AU - Hering, Bernhard J.

PY - 2010/4

Y1 - 2010/4

N2 - Background. Purified tissue dissociation enzymes (TDEs) are critical to successful human islet isolation required for clinical transplantation, but little is known about the characteristics of the key enzymes-class I (C1) and class II (C2) collagenase from Clostridium histolyticum-used in these procedures. Here, we show the differences between the C1 collagenase found in purified collagenase products manufactured by three suppliers and the impact of differences in C1 between two suppliers on human islet yield. Methods. Collagenase from Roche, Serva/Nordmark (Uetersen, Germany), and VitaCyte (Indianapolis, IN) were analyzed by analytical high-performance liquid chromatography and collagen degradation activity (CDA), an assay that preferentially detects intact C1 collagenase. Human islet isolations were performed using current standard practices. Results. These studies showed that the highest amount of intact C1 that correlated with a high specific CDA (CDA unit per milligram of protein). The highest specific CDA was found in VitaCyte product followed by the Roche and Serva/Nordmark products. The products of VitaCyte were used successfully for human islet isolation (n=14) with an average final islet yield obtained was 419,100±150,900 islet equivalent number (IEQ) (4147±1759 IEQ/g pancreas). Four of these preparations were used successfully in clinical transplantation procedures. These TDEs gave significantly better results when compared with earlier data where 27 isolations were performed using Serva NB1 collagenase and NB neutral protease where the final islet yield was 217,500±152,400 IEQ (2134±1524 IEQ/g pancreas). Conclusions. These data indicate the importance of intact C1 and the use of the appropriate analytical assays to correlate biochemical characteristics of TDEs to islet quality and yield.

AB - Background. Purified tissue dissociation enzymes (TDEs) are critical to successful human islet isolation required for clinical transplantation, but little is known about the characteristics of the key enzymes-class I (C1) and class II (C2) collagenase from Clostridium histolyticum-used in these procedures. Here, we show the differences between the C1 collagenase found in purified collagenase products manufactured by three suppliers and the impact of differences in C1 between two suppliers on human islet yield. Methods. Collagenase from Roche, Serva/Nordmark (Uetersen, Germany), and VitaCyte (Indianapolis, IN) were analyzed by analytical high-performance liquid chromatography and collagen degradation activity (CDA), an assay that preferentially detects intact C1 collagenase. Human islet isolations were performed using current standard practices. Results. These studies showed that the highest amount of intact C1 that correlated with a high specific CDA (CDA unit per milligram of protein). The highest specific CDA was found in VitaCyte product followed by the Roche and Serva/Nordmark products. The products of VitaCyte were used successfully for human islet isolation (n=14) with an average final islet yield obtained was 419,100±150,900 islet equivalent number (IEQ) (4147±1759 IEQ/g pancreas). Four of these preparations were used successfully in clinical transplantation procedures. These TDEs gave significantly better results when compared with earlier data where 27 isolations were performed using Serva NB1 collagenase and NB neutral protease where the final islet yield was 217,500±152,400 IEQ (2134±1524 IEQ/g pancreas). Conclusions. These data indicate the importance of intact C1 and the use of the appropriate analytical assays to correlate biochemical characteristics of TDEs to islet quality and yield.

KW - Collagenase

KW - Collagenase assay

KW - Islet isolation

KW - Islet transplantation

KW - Tissue Dissociation

UR - http://www.scopus.com/inward/record.url?scp=77951295605&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77951295605&partnerID=8YFLogxK

U2 - 10.1097/TP.0b013e3181d21e9a

DO - 10.1097/TP.0b013e3181d21e9a

M3 - Article

C2 - 20300051

AN - SCOPUS:77951295605

VL - 89

SP - 954

EP - 961

JO - Transplantation

JF - Transplantation

SN - 0041-1337

IS - 8

ER -