Gene Transfer of Constitutively Active Akt Markedly Improves Human Islet Transplant Outcomes in Diabetic Severe Combined Immunodeficient Mice

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FIG. 2.
FIG. 2.

Proliferation in human islet cells transduced with Adv-CA-Akt. A: [3H]Thymidine incorporation in isolated human islets transduced with CA-Akt and LacZ. One hundred human IEQs were harvested 24 h after transduction and incubated 48 h in RPMI medium without FBS. [3H]Thymidine was added during the last 24 h. [3H]Thymidine incorporation is expressed as a percentage of the incorporation in uninfected islets (100%). B: Representative photomicrographs showing BrdU (red) and insulin (green) staining in sections of uninfected human islets and human islets transduced with Adv-LacZ and Adv-CA-Akt at the indicated MOI. Islets were treated as described for the [3H]thymidine experiments except BrdU was added instead of [3H]thymidine. Arrowheads indicate BrdU-positive nuclei. Hoestch 33258 (blue) staining was performed to detect cell nuclei. The magnification used is ×400 except for the lower right panel in which BrdU-positive β-cell nuclei are shown, which was taken at ×1,000. C: Quantitation of BrdU incorporation into uninfected human islets and Adv-LacZ- and Adv-CA-Akt–transduced human islets at the indicated MOI. The minimum number of nuclei counted per section was 1,000. *P < 0.05 vs. uninfected and Adv-LacZ islets. D. Quantitation of BrdU-positive β-cells as a function of the total number of human β-cells in sections of uninfected human islets or Adv-LacZ–and Adv-CA-Akt–transduced human islets. For these experiments, an MOI of 500 was used. Results are means ± SE. *P < 0.05 vs. uninfected and Adv-LacZ–transduced islets. Six different human islet preparations were examined.

This Article

  1. Diabetes vol. 54 no. 6 1664-1675