Anatomy knowledge has been found to include both spatial and non-spatial components. However, no systematic evaluation of studies relating spatial abilities and anatomy knowledge has been undertaken. The objective of this study was to conduct a systematic review of the relationship between spatial abilities test and anatomy knowledge assessment. A literature search was done up to March 20, 2014 in Scopus and in several databases on the OvidSP and EBSCOhost platforms. Of the 556 citations obtained, 38 articles were identified and fully reviewed yielding 21 eligible articles and their quality were formally assessed. Non-significant relationships were found between spatial abilities test and anatomy knowledge assessment using essays and non-spatial multiple-choice questions. Significant relationships were observed between spatial abilities test and anatomy knowledge assessment using practical examination, three-dimensional synthesis from two-dimensional views, drawing of views, and cross-sections. Relationships between spatial abilities test and anatomy knowledge assessment using spatial multiple-choice questions were unclear. The results of this systematic review provide evidence for spatial and non-spatial methods of anatomy knowledge assessment.