Juvenile play shows sex differences in animals and humans. Animal studies and a recent study in humans suggest that testosterone exposure during early development plays a key role. Here we report on the relationship between children’s sex-typed play behavior and digit ratio (2D:4D), a putative negative correlate of prenatal testosterone. 2D:4D and sex-typed play behavior as assessed by parents were negatively correlated in a sample of 83 pre-school boys but not in a sample of 93 girls. This finding lends some support to the ideas that early testosterone has a masculinising effect upon sex-typed play behavior in humans and that 2D:4D is a valuable tool for studying effects of early testosterone on human behavior.