Tumorigenesis in glioblastoma multiforme (GBM) is complex and depends on interactions of tumor cells with the tumor microenvironment. A limiting factor in translating preclinical GBM treatment studies is being able to longitudinally and non-invasively map the tumor microenvironment in the same tumor over days and weeks. Here we describe a multi-modal MRI study of glioma growth and metabolism in human-derived models of GBM that differ in levels of hypoxia, angiogenesis, and necrosis. We demonstrate successful longitudinal mapping of tumor growth and extracellular pH in the same U87 and U251 gliomas. We find that despite significant tumor growth, acidosis plateaus early.