Abstract Background Current isolation techniques recover only 20% to 50% of the pancreatic islets. Brain death (BD) is characterized by activation of proinflammatory cytokines (PICs) with reduced islet yields and functionality. We previously reported that 17β-estradiol (E2) induces cytoprotection to human islets exposed to PICs. Furthermore, inhibition of PIC release has been demonstrated after E2 treatment. In the present study, we evaluated if E2 treatment to BD donors would improve pancreatic islet recovery and functionality. Methods BD was induced in male, 250- to 350-g Lewis rats by inflation of a Fogarty catheter placed intracranially. Rats were mechanically ventilated for 6 hours. Only rats with mean arterial blood pressure >75 mm Hg were used. Animals (n = 6) received E2 (1mg/kg/iv immediately after BD induction), vehicle (V), or the combination of 17β-estradiol and a selective estrogen receptor antagonist ICI 182,780 (ICI, 3mg/kg/ip/1 hour before BD induction). Islet viability was determined by ethidium bromide–acridine orange. PICs were assessed by ELISA. Islet functionality was determined by static incubation and glucose disposal rate (Kg) after intraportal transplantation (3000 islet equivalent[IEQ]/syngeneic streptozotocin-induced diabetic rat). Results A 2- to 3-fold reduction in TNF-α, IL-1β, and IL-6 was demonstrated in BD donors given E2; this effect reversed by ICI 182,780. Pancreatic sections from control BD donors presented 26.5% ± 4% TUNEL-positive β-cells compared with 15.1% ± 3% in 17β-estradio–treated animals. Islet recovery was enhanced in E2-treated donors (1233.4 ± 123 IEQ/pancreas) compared with controls (725 ± 224 IEQ, P < .05). Islet viability was significantly enhanced by E2. Higher islet functionality was demonstrated in vitro and in vivo after transplantation in islets recovered from E2-treated BD donors. Conclusions Islet recovery and functionality in vitro and in vivo were significantly improved by 17β-estradiol treatment to BD donors. These observations may lead to strategies to reduce the effects of BD on isolated islets and improve the results in clinical islet transplantation.