| Summary: | Beta-cell survival is low following islet transplantation and this may be linked to a delay in revascularisation of donor cells. This decrease in oxygen supply is termed hypoxia, the result of which is detrimental to beta-cell survival. The current research sought to investigate post-transplant beta-cell viability and function by investigating the effects of low oxygen levels on MIN6 pancreatic beta-cells. MIN6 beta-cells were exposed to 1% oxygen (hypoxia) or 21% oxygen (normoxia) over a period of 72 hours. Viability was assessed by MTT assay and cell number was determined by haemocytometer count at 0 hours (normoxia), 24 hours, 48 hours and 72 hours. A Hoechst propidium iodide (HPI) stain was used to identify beta-cell apoptosis or necrosis during hypoxia. Western blot analyses were performed to determine the PI3K, pAkt, pAMPK, PDX-1, GLUT2 and pS6K protein levels. Real time PCR was used to estimate glut2, vegf, hif and insulin gene expression by MIN6 cells following exposure to hypoxia over various time points.
|
|---|
