Transplantation of macroencapsulated tissue-engineered grafts (TEGs) is being investigated as a treatment for type 1 diabetes, but there is a critical need to measure TEG viability both in vitro and in vivo. Oxygen deficiency is the most critical issue preventing widespread implementation of TEG transplantation and delivery of supplemental oxygen (DSO) has been shown to enhance TEG survival and function in vivo. In this study, we demonstrate the first use of oxygen-17 magnetic resonance spectroscopy (O-17-MRS) to measure the oxygen consumption rate (OCR) of TEGs and show that in addition to providing therapeutic benefits to TEGs, DSO with O-17(2) can also enable measurements of TEG viability. Macroencapsulated TEGs containing beta TC3 murine insulinoma cells were prepared with three fractional viabilities and provided with O-17(2). Cellular metabolism of O-17(2) into nascent mitochondrial water (H-2 O-17) was monitored by O-17-MRS and, from the measured data, OCR was calculated. For comparison, OCR was simultaneously measured on a separate, but equivalent sample of cells with a well-established stirred microchamber technique. OCR measured by O-17-MRS agreed well with measurements made in the stirred microchamber device. These studies confirm that O-17-MRS can quantify TEG viability noninvasively. (C) 2016 Wiley Periodicals, Inc.