Adenovirus mediated gene transduction of primarily isolated mouse islets

Michiki Narushima, Teru Okitsu, Atsushi Miki, Chen Yong, Kazuya Kobayashi, Yukihide Yonekawa, Kimiaki Tanaka, Hideaki Ikeda, Shinichi Matsumoto, Noriaki Tanaka, Naoya Kobayashi

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Expansion of pancreatic islet cell populations, especially the beta cells, using a currently available ex vivo gene transfer technology is important to develop cell therapies to treat Type I diabetes. In this study, we evaluated adenovirus mediated gene transfer efficiency in primarily isolated mouse islet cells in two types of culture conditions: freshly isolated suspended islets and cultured islets with monolayer formation. A recombinant replication deficient adenovirus vector encoding a green fluorescence protein (CFP) cDNA, Ad/ CMV-GFP, was used in the present transduction experiments. Rat 804G derived extracellular matrix (804G-ECM) and 3-isobutyl-1-methylxanthine (IBMX) were used to facilitate monolayer formation of the isolated mouse pancreatic islets. Suspended islets were transfected with Ad/CMV-GFP at more than 95% efficiency. However, analysis of immunohistochemical stains for insulin and glucagon in thin sliced sections of the islets revealed that GFP expression was localized just in the outer cells of the islets, almost all of which were glucagon positive alpha-cells. The beta-cells existing at the inner area of the suspended islets were CFP negative. In contrast, under the condition of the islets in monolayer formation cultures, all of the islet cells including the beta-cells were efficiently infected with Ad/CMV-GFP. To achieve an efficient adenoviral gene transfer to the pancreatic beta-cells, monolayer formation of the islets is critical. Such culture conditions were facilitated by combining 804G-ECM with IBMX.

Original languageEnglish (US)
Pages (from-to)586-590
Number of pages5
JournalASAIO Journal
Volume50
Issue number6
DOIs
StatePublished - Nov 2004
Externally publishedYes

Fingerprint

Gene transfer
Islets of Langerhans
Adenoviridae
Monolayers
Genes
Glucagon
1-Methyl-3-isobutylxanthine
Extracellular Matrix
Insulin
Medical problems
Technology Transfer
Rats
Coloring Agents
Complementary DNA
Fluorescence
Cells
Insulin-Secreting Cells
Cell- and Tissue-Based Therapy
Proteins
Type 1 Diabetes Mellitus

ASJC Scopus subject areas

  • Biophysics
  • Bioengineering

Cite this

Narushima, M., Okitsu, T., Miki, A., Yong, C., Kobayashi, K., Yonekawa, Y., ... Kobayashi, N. (2004). Adenovirus mediated gene transduction of primarily isolated mouse islets. ASAIO Journal, 50(6), 586-590. https://doi.org/10.1097/01.MAT.0000142877.18621.BC

Adenovirus mediated gene transduction of primarily isolated mouse islets. / Narushima, Michiki; Okitsu, Teru; Miki, Atsushi; Yong, Chen; Kobayashi, Kazuya; Yonekawa, Yukihide; Tanaka, Kimiaki; Ikeda, Hideaki; Matsumoto, Shinichi; Tanaka, Noriaki; Kobayashi, Naoya.

In: ASAIO Journal, Vol. 50, No. 6, 11.2004, p. 586-590.

Research output: Contribution to journalArticle

Narushima, M, Okitsu, T, Miki, A, Yong, C, Kobayashi, K, Yonekawa, Y, Tanaka, K, Ikeda, H, Matsumoto, S, Tanaka, N & Kobayashi, N 2004, 'Adenovirus mediated gene transduction of primarily isolated mouse islets', ASAIO Journal, vol. 50, no. 6, pp. 586-590. https://doi.org/10.1097/01.MAT.0000142877.18621.BC
Narushima M, Okitsu T, Miki A, Yong C, Kobayashi K, Yonekawa Y et al. Adenovirus mediated gene transduction of primarily isolated mouse islets. ASAIO Journal. 2004 Nov;50(6):586-590. https://doi.org/10.1097/01.MAT.0000142877.18621.BC
Narushima, Michiki ; Okitsu, Teru ; Miki, Atsushi ; Yong, Chen ; Kobayashi, Kazuya ; Yonekawa, Yukihide ; Tanaka, Kimiaki ; Ikeda, Hideaki ; Matsumoto, Shinichi ; Tanaka, Noriaki ; Kobayashi, Naoya. / Adenovirus mediated gene transduction of primarily isolated mouse islets. In: ASAIO Journal. 2004 ; Vol. 50, No. 6. pp. 586-590.
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