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

(Downloading may take up to 30 seconds. If the slide opens in your browser, select File -> Save As to save it.)

Click on image to view larger version.

FIG. 3.FIG. 3.
FIG. 3.

Quantitation of cell death in human islets after glucose and serum deprivation (A and B) and chronic exposure to 22 mmol/l glucose for 5 days (C and D). A: Representative photomicrographs of sections of uninfected human islets and Adv-LacZ–and Adv-CA-Akt–transduced human islets incubated in RPMI medium devoid of glucose and serum and stained for insulin (red) and TUNEL (green). Twenty-four hours after transduction, islets were placed in RPMI containing 5 mmol/l glucose and 10% FBS or RPMI medium devoid of glucose and FBS for 24 h. Arrowheads indicate TUNEL-positive nuclei. As control, a section of Adv-LacZ–transduced human islets incubated in complete conditioned RPMI medium (5 mmol/l glucose and 10% FBS) is shown. B: Quantitation of TUNEL-positive β-cell (insulin-positive) nuclei in these sections of uninfected human islets and Adv-LacZ–or Adv-CA-Akt–transduced human islets incubated in conditioned medium or medium devoid of glucose and FBS (see above). C: Representative photomicrographs of sections of uninfected human islets and Adv-LacZ–and Adv-CA-Akt–transduced human islets incubated in RPMI medium containing 22 mmol/l glucose for 5 days and stained for insulin (red) and TUNEL (green). Twenty-four hours after transduction, islets were placed in RPMI containing 5 mmol/l glucose and 10% FBS or RPMI containing 22 mmol/l glucose and 10% FBS for 5 days. Arrowheads indicate TUNEL-positive nuclei. As control, a section of Adv-LacZ–transduced human islets incubated in complete conditioned RPMI medium (5 mmol/l glucose and 10% FBS) is shown. D: Quantitation of TUNEL-positive β-cell nuclei in these sections of uninfected human islets and Adv-LacZ–or Adv-CA-Akt–transduced human islets incubated in medium containing either 5 or 22 mmol/l glucose (see above). For these experiments, an MOI of 500 was used. Hoestch 33258 (blue) staining was performed to detect cell nuclei. Results are means ± SE. *P < 0.05 vs. uninfected and Adv-LacZ–transduced treated islets. Five different human islet preparations were examined.

This Article

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