Ischemic tissue damage resulting from the surgical extraction procedure of donor kidneys remains an important clinical problem in kidney transplantation. This thesis aims to gain insight and find potential solutions to mitigate the impact of ischemic injury. In addition to quantification of the impact of the duration of ischemia, also the role of temperature during this ischemic period was investigated. Furthermore, the effect of temperature on mitochondrial function and reactive oxygen species production were determined and new strategies to decrease ischemic injury were evaluated. These strategies include the use of hydrogen sulphide, oxygen and glucose during the flush-out at the start of extraction procedure. Hydrogen sulphide showed interesting capacities in inducing a hypometabolic state and reducing inflammatory markers but failed to prevent ischemic injury. The use of oxygen and glucose improved the energy status of the kidneys after the flush-out. Lastly, a new way of visualizing renal cortical microperfusion was studied, thereby improving the assessment of damage resulting from ischemia and getting better insight into organ quality.