Improved in vivo pancreatic islet function after prolonged in vitro islet culture

A. Osama Gaber, Daniel W. Fraga, Christopher S. Callicutt, Ivan Gerling, Omaima M. Sabek, Malak Y. Kotb

Research output: Contribution to journalArticle

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Abstract

Background. Difficulties with recovering and preserving pancreatic islets have hampered progress in islet transplantation. In previous in vitro studies, our laboratory successfully demonstrated that using serum-free medium for prolonged pancreatic islet culture allows postculture recovery ratios greater than those obtained with standard media with sustained in vitro islet function. The goal of this study was to determine whether culturing of islets in a modified serum-free medium (M-SFM) would sustain function in vivo. Methods. Islets were isolated from pancreata procured from 12 cadaveric organ donors and cultured in the M-SFM for up to 2 months, cryopreserved at -70°C within 1-3 days of isolation for 2 months, or placed in short-term culture (3-5 days) before their transplantation under the kidney capsule of nonobese diabetic-severe combined immunodeficient mice (n=4-7 per group/time point). In vivo islet function was assessed by measuring the production of human insulin and C-peptide over a period of 3-15 months. Results. After extended culture of islets in M-SFM for 1 or 2 months, transplanted islets maintained their viability, and in some instances in vivo function improved when compared with short-term cultured islets transplanted from the same preparation (P<0.01). Improvement was particularly evident for islets cultured for 1 month. Furthermore, when compared with cryopreserved preparations, early function (postoperative day 7) of islets from 1-month culture preparations was statistically better (P<0.05). Prolonged culture in M-SFM had no significant impact on long-term function, inasmuch as cultured islets functioned for more than 120 days. Conclusion. Thess data demonstrate that prolonged islet culture in M-SFM sustained viability and function, and in some instances had a positive effect on in vivo islet function, particularly in the 1-month cultures. No negative effect on long-term in vivo function was demonstrated in this study. Confirmation in clinical models utilizing extended (1-2 months) islet culture in M-SFM could significantly enhance islet transplantation by allowing the identification of best-matched recipients, pretransplantation recipient conditioning, and possible pretransplantation islet modifications to promote engraftment and prolonged graft function.

Original languageEnglish (US)
Pages (from-to)1730-1736
Number of pages7
JournalTransplantation
Volume72
Issue number11
DOIs
StatePublished - Dec 15 2001
Externally publishedYes

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Serum-Free Culture Media
Islets of Langerhans
Islets of Langerhans Transplantation
SCID Mice
C-Peptide
In Vitro Techniques
Kidney Transplantation
Capsules
Pancreas
Tissue Donors
Insulin
Transplants

All Science Journal Classification (ASJC) codes

  • Transplantation

Cite this

Gaber, A. O., Fraga, D. W., Callicutt, C. S., Gerling, I., Sabek, O. M., & Kotb, M. Y. (2001). Improved in vivo pancreatic islet function after prolonged in vitro islet culture. Transplantation, 72(11), 1730-1736. https://doi.org/10.1097/00007890-200112150-00005

Improved in vivo pancreatic islet function after prolonged in vitro islet culture. / Gaber, A. Osama; Fraga, Daniel W.; Callicutt, Christopher S.; Gerling, Ivan; Sabek, Omaima M.; Kotb, Malak Y.

In: Transplantation, Vol. 72, No. 11, 15.12.2001, p. 1730-1736.

Research output: Contribution to journalArticle

Gaber, A. Osama ; Fraga, Daniel W. ; Callicutt, Christopher S. ; Gerling, Ivan ; Sabek, Omaima M. ; Kotb, Malak Y. / Improved in vivo pancreatic islet function after prolonged in vitro islet culture. In: Transplantation. 2001 ; Vol. 72, No. 11. pp. 1730-1736.
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T1 - Improved in vivo pancreatic islet function after prolonged in vitro islet culture

AU - Gaber, A. Osama

AU - Fraga, Daniel W.

AU - Callicutt, Christopher S.

AU - Gerling, Ivan

AU - Sabek, Omaima M.

AU - Kotb, Malak Y.

PY - 2001/12/15

Y1 - 2001/12/15

N2 - Background. Difficulties with recovering and preserving pancreatic islets have hampered progress in islet transplantation. In previous in vitro studies, our laboratory successfully demonstrated that using serum-free medium for prolonged pancreatic islet culture allows postculture recovery ratios greater than those obtained with standard media with sustained in vitro islet function. The goal of this study was to determine whether culturing of islets in a modified serum-free medium (M-SFM) would sustain function in vivo. Methods. Islets were isolated from pancreata procured from 12 cadaveric organ donors and cultured in the M-SFM for up to 2 months, cryopreserved at -70°C within 1-3 days of isolation for 2 months, or placed in short-term culture (3-5 days) before their transplantation under the kidney capsule of nonobese diabetic-severe combined immunodeficient mice (n=4-7 per group/time point). In vivo islet function was assessed by measuring the production of human insulin and C-peptide over a period of 3-15 months. Results. After extended culture of islets in M-SFM for 1 or 2 months, transplanted islets maintained their viability, and in some instances in vivo function improved when compared with short-term cultured islets transplanted from the same preparation (P<0.01). Improvement was particularly evident for islets cultured for 1 month. Furthermore, when compared with cryopreserved preparations, early function (postoperative day 7) of islets from 1-month culture preparations was statistically better (P<0.05). Prolonged culture in M-SFM had no significant impact on long-term function, inasmuch as cultured islets functioned for more than 120 days. Conclusion. Thess data demonstrate that prolonged islet culture in M-SFM sustained viability and function, and in some instances had a positive effect on in vivo islet function, particularly in the 1-month cultures. No negative effect on long-term in vivo function was demonstrated in this study. Confirmation in clinical models utilizing extended (1-2 months) islet culture in M-SFM could significantly enhance islet transplantation by allowing the identification of best-matched recipients, pretransplantation recipient conditioning, and possible pretransplantation islet modifications to promote engraftment and prolonged graft function.

AB - Background. Difficulties with recovering and preserving pancreatic islets have hampered progress in islet transplantation. In previous in vitro studies, our laboratory successfully demonstrated that using serum-free medium for prolonged pancreatic islet culture allows postculture recovery ratios greater than those obtained with standard media with sustained in vitro islet function. The goal of this study was to determine whether culturing of islets in a modified serum-free medium (M-SFM) would sustain function in vivo. Methods. Islets were isolated from pancreata procured from 12 cadaveric organ donors and cultured in the M-SFM for up to 2 months, cryopreserved at -70°C within 1-3 days of isolation for 2 months, or placed in short-term culture (3-5 days) before their transplantation under the kidney capsule of nonobese diabetic-severe combined immunodeficient mice (n=4-7 per group/time point). In vivo islet function was assessed by measuring the production of human insulin and C-peptide over a period of 3-15 months. Results. After extended culture of islets in M-SFM for 1 or 2 months, transplanted islets maintained their viability, and in some instances in vivo function improved when compared with short-term cultured islets transplanted from the same preparation (P<0.01). Improvement was particularly evident for islets cultured for 1 month. Furthermore, when compared with cryopreserved preparations, early function (postoperative day 7) of islets from 1-month culture preparations was statistically better (P<0.05). Prolonged culture in M-SFM had no significant impact on long-term function, inasmuch as cultured islets functioned for more than 120 days. Conclusion. Thess data demonstrate that prolonged islet culture in M-SFM sustained viability and function, and in some instances had a positive effect on in vivo islet function, particularly in the 1-month cultures. No negative effect on long-term in vivo function was demonstrated in this study. Confirmation in clinical models utilizing extended (1-2 months) islet culture in M-SFM could significantly enhance islet transplantation by allowing the identification of best-matched recipients, pretransplantation recipient conditioning, and possible pretransplantation islet modifications to promote engraftment and prolonged graft function.

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