Development of an automated computer-controlled islet isolation system

Jonathan R T Lakey, Garth L. Warnock, Mark Brierton, Ziliang Ao, Bernhard J. Hering, Nick J M London, Camillo Ricordi, Frank Corbin, Ray V. Rajotte

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Before clinical islet transplantation can become an effective and reliable treatment for type 1 diabetic patients, there must be significant improvements in the methods employed for the isolation of islets of Langerhans. We have developed an automated cell extraction system (ACES), which allows computer control of the isolation process. As well, it incorporates a novel method of recombining dissociated pancreatic tissue. Following initial system design and testing to determine the optimal system configuration, a series of 12 consecutive canine islet isolations were performed. Pancreases were perfused with collagenase via the duct and dissociated and recombined using either the standard Ricordi-based protocol (group 1, n = 6) or dissociated and recombined using the ACES system (group 2, n = 6). A total of 90.8 ± 21 x 103 islet equivalents (IE) (mean ± SEM) were recovered in group 1 vs. 99 ± 14 x 103 IE in group 2 (p = NS, student unpaired t-test). Following Ficoll purification the recovery was 56.2 ± 14 x 103 IE for group 1 vs. 54.7 ± 11 x 103 IE for group 2 (p = NS). Viability was equivalent with an 8.6-fold increase in insulin secretion for group 1 and an 8.8-fold increase for group 2 when the islets were exposed to high glucose solution supplemented with IBMX (3-isobutyl-1-methylxanthine) during static incubation. In vivo function was equivalent following transplantation of 2000 IE under the kidney capsule of alloxaninduced diabetic nude mice with five of six and five of seven mice surviving long-term (>50 days posttransplant) (groups 1 and 2, respectively). This data shows that an entirely automated pancreatic islet extraction system can result in effective canine islet recovery without compromising islet yields and viability. The ACES system has several advantages over the standard isolation protocol. These include: 1) computer control and monitoring over all phases of the isolation, 2) a single-use sterile disposable tubing set, and 3) a novel method of tissue recombination.

Original languageEnglish
Pages (from-to)47-57
Number of pages11
JournalCell Transplantation
Volume6
Issue number1
DOIs
StatePublished - Jan 1 1997

Fingerprint

Islets of Langerhans Transplantation
Islets of Langerhans
Canidae
Computer control
1-Methyl-3-isobutylxanthine
Ficoll
Collagenases
Tissue
Nude Mice
Recovery
Optimal systems
Genetic Recombination
Capsules
Pancreas
Insulin
Tubing
Ducts
Students
Purification
Glucose

Keywords

  • Diabetes
  • Islet isolation
  • Transplantation

ASJC Scopus subject areas

  • Cell Biology
  • Transplantation

Cite this

Lakey, J. R. T., Warnock, G. L., Brierton, M., Ao, Z., Hering, B. J., London, N. J. M., ... Rajotte, R. V. (1997). Development of an automated computer-controlled islet isolation system. Cell Transplantation, 6(1), 47-57. https://doi.org/10.1016/S0963-6897(96)00137-6

Development of an automated computer-controlled islet isolation system. / Lakey, Jonathan R T; Warnock, Garth L.; Brierton, Mark; Ao, Ziliang; Hering, Bernhard J.; London, Nick J M; Ricordi, Camillo; Corbin, Frank; Rajotte, Ray V.

In: Cell Transplantation, Vol. 6, No. 1, 01.01.1997, p. 47-57.

Research output: Contribution to journalArticle

Lakey, JRT, Warnock, GL, Brierton, M, Ao, Z, Hering, BJ, London, NJM, Ricordi, C, Corbin, F & Rajotte, RV 1997, 'Development of an automated computer-controlled islet isolation system', Cell Transplantation, vol. 6, no. 1, pp. 47-57. https://doi.org/10.1016/S0963-6897(96)00137-6
Lakey JRT, Warnock GL, Brierton M, Ao Z, Hering BJ, London NJM et al. Development of an automated computer-controlled islet isolation system. Cell Transplantation. 1997 Jan 1;6(1):47-57. https://doi.org/10.1016/S0963-6897(96)00137-6
Lakey, Jonathan R T ; Warnock, Garth L. ; Brierton, Mark ; Ao, Ziliang ; Hering, Bernhard J. ; London, Nick J M ; Ricordi, Camillo ; Corbin, Frank ; Rajotte, Ray V. / Development of an automated computer-controlled islet isolation system. In: Cell Transplantation. 1997 ; Vol. 6, No. 1. pp. 47-57.
@article{f30b614350b14630998e605d43139eb2,
title = "Development of an automated computer-controlled islet isolation system",
abstract = "Before clinical islet transplantation can become an effective and reliable treatment for type 1 diabetic patients, there must be significant improvements in the methods employed for the isolation of islets of Langerhans. We have developed an automated cell extraction system (ACES), which allows computer control of the isolation process. As well, it incorporates a novel method of recombining dissociated pancreatic tissue. Following initial system design and testing to determine the optimal system configuration, a series of 12 consecutive canine islet isolations were performed. Pancreases were perfused with collagenase via the duct and dissociated and recombined using either the standard Ricordi-based protocol (group 1, n = 6) or dissociated and recombined using the ACES system (group 2, n = 6). A total of 90.8 ± 21 x 103 islet equivalents (IE) (mean ± SEM) were recovered in group 1 vs. 99 ± 14 x 103 IE in group 2 (p = NS, student unpaired t-test). Following Ficoll purification the recovery was 56.2 ± 14 x 103 IE for group 1 vs. 54.7 ± 11 x 103 IE for group 2 (p = NS). Viability was equivalent with an 8.6-fold increase in insulin secretion for group 1 and an 8.8-fold increase for group 2 when the islets were exposed to high glucose solution supplemented with IBMX (3-isobutyl-1-methylxanthine) during static incubation. In vivo function was equivalent following transplantation of 2000 IE under the kidney capsule of alloxaninduced diabetic nude mice with five of six and five of seven mice surviving long-term (>50 days posttransplant) (groups 1 and 2, respectively). This data shows that an entirely automated pancreatic islet extraction system can result in effective canine islet recovery without compromising islet yields and viability. The ACES system has several advantages over the standard isolation protocol. These include: 1) computer control and monitoring over all phases of the isolation, 2) a single-use sterile disposable tubing set, and 3) a novel method of tissue recombination.",
keywords = "Diabetes, Islet isolation, Transplantation",
author = "Lakey, {Jonathan R T} and Warnock, {Garth L.} and Mark Brierton and Ziliang Ao and Hering, {Bernhard J.} and London, {Nick J M} and Camillo Ricordi and Frank Corbin and Rajotte, {Ray V.}",
year = "1997",
month = "1",
day = "1",
doi = "10.1016/S0963-6897(96)00137-6",
language = "English",
volume = "6",
pages = "47--57",
journal = "Cell Transplantation",
issn = "0963-6897",
publisher = "Cognizant Communication Corporation",
number = "1",

}

TY - JOUR

T1 - Development of an automated computer-controlled islet isolation system

AU - Lakey, Jonathan R T

AU - Warnock, Garth L.

AU - Brierton, Mark

AU - Ao, Ziliang

AU - Hering, Bernhard J.

AU - London, Nick J M

AU - Ricordi, Camillo

AU - Corbin, Frank

AU - Rajotte, Ray V.

PY - 1997/1/1

Y1 - 1997/1/1

N2 - Before clinical islet transplantation can become an effective and reliable treatment for type 1 diabetic patients, there must be significant improvements in the methods employed for the isolation of islets of Langerhans. We have developed an automated cell extraction system (ACES), which allows computer control of the isolation process. As well, it incorporates a novel method of recombining dissociated pancreatic tissue. Following initial system design and testing to determine the optimal system configuration, a series of 12 consecutive canine islet isolations were performed. Pancreases were perfused with collagenase via the duct and dissociated and recombined using either the standard Ricordi-based protocol (group 1, n = 6) or dissociated and recombined using the ACES system (group 2, n = 6). A total of 90.8 ± 21 x 103 islet equivalents (IE) (mean ± SEM) were recovered in group 1 vs. 99 ± 14 x 103 IE in group 2 (p = NS, student unpaired t-test). Following Ficoll purification the recovery was 56.2 ± 14 x 103 IE for group 1 vs. 54.7 ± 11 x 103 IE for group 2 (p = NS). Viability was equivalent with an 8.6-fold increase in insulin secretion for group 1 and an 8.8-fold increase for group 2 when the islets were exposed to high glucose solution supplemented with IBMX (3-isobutyl-1-methylxanthine) during static incubation. In vivo function was equivalent following transplantation of 2000 IE under the kidney capsule of alloxaninduced diabetic nude mice with five of six and five of seven mice surviving long-term (>50 days posttransplant) (groups 1 and 2, respectively). This data shows that an entirely automated pancreatic islet extraction system can result in effective canine islet recovery without compromising islet yields and viability. The ACES system has several advantages over the standard isolation protocol. These include: 1) computer control and monitoring over all phases of the isolation, 2) a single-use sterile disposable tubing set, and 3) a novel method of tissue recombination.

AB - Before clinical islet transplantation can become an effective and reliable treatment for type 1 diabetic patients, there must be significant improvements in the methods employed for the isolation of islets of Langerhans. We have developed an automated cell extraction system (ACES), which allows computer control of the isolation process. As well, it incorporates a novel method of recombining dissociated pancreatic tissue. Following initial system design and testing to determine the optimal system configuration, a series of 12 consecutive canine islet isolations were performed. Pancreases were perfused with collagenase via the duct and dissociated and recombined using either the standard Ricordi-based protocol (group 1, n = 6) or dissociated and recombined using the ACES system (group 2, n = 6). A total of 90.8 ± 21 x 103 islet equivalents (IE) (mean ± SEM) were recovered in group 1 vs. 99 ± 14 x 103 IE in group 2 (p = NS, student unpaired t-test). Following Ficoll purification the recovery was 56.2 ± 14 x 103 IE for group 1 vs. 54.7 ± 11 x 103 IE for group 2 (p = NS). Viability was equivalent with an 8.6-fold increase in insulin secretion for group 1 and an 8.8-fold increase for group 2 when the islets were exposed to high glucose solution supplemented with IBMX (3-isobutyl-1-methylxanthine) during static incubation. In vivo function was equivalent following transplantation of 2000 IE under the kidney capsule of alloxaninduced diabetic nude mice with five of six and five of seven mice surviving long-term (>50 days posttransplant) (groups 1 and 2, respectively). This data shows that an entirely automated pancreatic islet extraction system can result in effective canine islet recovery without compromising islet yields and viability. The ACES system has several advantages over the standard isolation protocol. These include: 1) computer control and monitoring over all phases of the isolation, 2) a single-use sterile disposable tubing set, and 3) a novel method of tissue recombination.

KW - Diabetes

KW - Islet isolation

KW - Transplantation

UR - http://www.scopus.com/inward/record.url?scp=0031035567&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031035567&partnerID=8YFLogxK

U2 - 10.1016/S0963-6897(96)00137-6

DO - 10.1016/S0963-6897(96)00137-6

M3 - Article

C2 - 9040955

AN - SCOPUS:0031035567

VL - 6

SP - 47

EP - 57

JO - Cell Transplantation

JF - Cell Transplantation

SN - 0963-6897

IS - 1

ER -