There is increasing evidence that biological sex contributes to the pathophysiological progression of diabetes and its associated complications. Specifically, males have a higher diabetic incidence, when compared to females of the same age and body mass index. The relative protection against diabetes corresponding to the female sex is ablated after menopause, suggesting a potential role for female sex hormones in preventing pancreatic dysfunction and subsequently, diabetes onset. Emerging in vivo and in vitro evidence showcases the potential for female sex hormone, 17β-Estradiol, to protect pancreatic β cells by regulating endoplasmic reticulum (ER) capacity. The ER plays a crucial role in the pancreatic cellular stress response to diabetes via the activation of the unfolded protein response (UPR). This study aims to delineate the mechanisms by which 17β-Estradiol promotes pancreatic health, by quantifying expression of adaptive and apoptotic UPR markers in both pancreatic β cell lines and pancreatic islets isolated from the ApoE-/-:Ins2+/Akita mouse model. Through the use of real-time PCR and immunofluorescence, we were able to characterize the potential of 17β-Estradiol to selectively mediate the UPR by differentially regulating its adaptive and apoptotic branches. The result of this study provides a better understanding of how sex plays a role in protecting pancreatic function in cell and murine models of diabetes. This is a crucial area in diabetes research that has vast potential applications, including the development of sex-dependent therapies to treat diabetes and its associated complications.