Changing circumstances alter the costs and benefits of choosing different mates and are thought to be reflected in women’s mate preferences. Indeed, several lines of reasoning, and some prior studies, suggest that individual differences in women’s preferences for cues of men’s underlying health will be more apparent among partnered women than among unpartnered women. The current study shows that preferences for male faces with masculine shape cues, characteristics that are thought to signal men’s underlying health, are positively correlated with partnered, but not unpartnered, women’s reported reproductive ambition (i.e., their desire to become pregnant). These findings (1) present new evidence for systematic variation in women’s mating strategies, (2) suggest that partnership status may be important for potentially adaptive variation in women’s mate preferences, and (3) suggest that reproductive ambition may influence women’s mate preferences. Alternative explanations for these findings, focusing on the possible effects of a range of variables that may be correlated with reproductive ambition in partnered women and influence their masculinity preferences, are also discussed.