Maintenance of glucose-sensitive insulin secretion of cryopreserved human islets with University of Wisconsin solution and ascorbic acid-2 glucoside

Takashi Arata, Teru Okitsu, Takuya Fukazawa, Hideaki Ikeda, Kazuya Kobayashi, Chen Yong, Yoshikazu Kosaka, Michiki Narushima, Junji Matsuoka, Itaru Yamamoto, Noriaki Tanaka, Jonathan R T Lakey, Naoya Kobayashi

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Normal human islet cells are an ideal source for pancreas-targeted cell therapies, but the availability of human donor pancreata for islet isolation is severely limited. To effectively utilize such scarce donor organs for cell therapies, it is crucial to develop an excellent isolation, effective cryopreservation, and efficient gene transfer techniques for the transportation of isolated cells. In the present study, we investigate the effect of University of Wisconsin (UW) solution and ascorbic acid-2 glucoside (AA2G) on the cryopreservation of human islets. We also evaluate the gene transfer efficiency of a lentiviral vector expressing the E. coli LacZ gene, Lt-NLS/LacZ, in human islets. Human islets were isolated with a standard digestion method at the University of Alberta. Isolated islets were transported to Japan for 40 h and then subjected to cryopreservation experiments. The following preservation solutions were tested: UW solution with 100 μg/mL of AA2G, UW solution, 100% fetal bovine serum (FBS), and CMRL supplemented with 10% FBS. Following three months of cryopreservation, the islets were thawed and analyzed for viability, glucose-sensitive insulin secretion, proinsulin gene expression profile, and in vivo engraftment. The islets were also subjected to monolayer formation with 804G-cell-line-derived extracellular matrix (ECM), followed by Lt-NLS/LacZ transduction. The viability, morphology, glucose-sensitive insulin secretion, proinsulin gene expression, and monolayer formation efficiency of the thawed cryopreserved islets are significantly better maintained by the use of UW solution. When AA2G (100 μg/mL) is combined with UW, such parameters are further improved. The adequate engraftment of UW + AA2G-cryopreserved human islets is achieved in the liver of nude mice. Efficient Lt-NLS/LacZ transduction is identified in monolayered islets cryopreserved with UW solution with AA2G. The present work demonstrates that the combination of UW solution with AA2G (100 μg/mL) would be a useful cryopreservation means for human islets. Human islets monolayer-cultured with 804G-derived ECM are efficiently transduced with a lentiviral vector Lt-NLS/LacZ.

Original languageEnglish
Pages (from-to)529-536
Number of pages8
JournalArtificial Organs
Volume28
Issue number6
DOIs
Publication statusPublished - Jun 2004

Fingerprint

Ascorbic acid
Insulin
Glucose
Maintenance
Cryopreservation
Gene transfer
Proinsulin
Monolayers
Gene expression
Cell- and Tissue-Based Therapy
Extracellular Matrix
Pancreas
Tissue Donors
Gene Transfer Techniques
Alberta
Lac Operon
ascorbic acid 2-O-glucoside
Glucosides
Liver
Escherichia coli

Keywords

  • Ascorbic acid-2 glucoside
  • Cryopreservation
  • Islets
  • Stimulation index
  • University of Wisconsin solution

ASJC Scopus subject areas

  • Biophysics

Cite this

Maintenance of glucose-sensitive insulin secretion of cryopreserved human islets with University of Wisconsin solution and ascorbic acid-2 glucoside. / Arata, Takashi; Okitsu, Teru; Fukazawa, Takuya; Ikeda, Hideaki; Kobayashi, Kazuya; Yong, Chen; Kosaka, Yoshikazu; Narushima, Michiki; Matsuoka, Junji; Yamamoto, Itaru; Tanaka, Noriaki; Lakey, Jonathan R T; Kobayashi, Naoya.

In: Artificial Organs, Vol. 28, No. 6, 06.2004, p. 529-536.

Research output: Contribution to journalArticle

Arata, T, Okitsu, T, Fukazawa, T, Ikeda, H, Kobayashi, K, Yong, C, Kosaka, Y, Narushima, M, Matsuoka, J, Yamamoto, I, Tanaka, N, Lakey, JRT & Kobayashi, N 2004, 'Maintenance of glucose-sensitive insulin secretion of cryopreserved human islets with University of Wisconsin solution and ascorbic acid-2 glucoside', Artificial Organs, vol. 28, no. 6, pp. 529-536. https://doi.org/10.1111/j.1525-1594.2004.07296.x
Arata, Takashi ; Okitsu, Teru ; Fukazawa, Takuya ; Ikeda, Hideaki ; Kobayashi, Kazuya ; Yong, Chen ; Kosaka, Yoshikazu ; Narushima, Michiki ; Matsuoka, Junji ; Yamamoto, Itaru ; Tanaka, Noriaki ; Lakey, Jonathan R T ; Kobayashi, Naoya. / Maintenance of glucose-sensitive insulin secretion of cryopreserved human islets with University of Wisconsin solution and ascorbic acid-2 glucoside. In: Artificial Organs. 2004 ; Vol. 28, No. 6. pp. 529-536.
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