With an increasing population living at a high altitude (HA), the impact of HA residence on human cognitive function has raised concerns. We recruited two groups of college students with one group born and grew up at HA until early adulthood and the control group born and grew up at near sea level (SL); the two groups were matched at age, gender ratio, educational level, the ancestral lines, and peripheral physiology (especially the hemoglobin concentration). A 2-back spatial working memory task was performed by each subject in the scanner while fMRI data were acquired. Compared to the SL control group, the HA group showed equal response accuracy, with more variance in reaction time and a larger average value. fMRI data indicated that both groups showed common activation patterns in the neural pathway typically associated with working memory. The HA group had greater activation at the left pyramis, the left superior temporal gyrus and less activation at the left middle occipital gyrus. Significant correlations were found within each group between the reaction times and BOLD signal change amplitudes at the frontal cortex and the precentral cortex. (Contains 4 figures and 1 table.)