Purpose: Women make essential contributions to the workforce in the United States but still hold a minority of senior leadership and chief executive officer positions in healthcare. The goal of this explanatory quantitative survey study was to measure the extent to which there are differences in the motivation to attain leadership positions between men and women. Methods: A stratified, random sample of 219 men and women employed in the healthcare industry were surveyed using the Motivation to Lead (MtL) questionnaire, a validated survey model that measures three leadership motivational factors. The data was analyzed using quantitative analysis. Principal Findings: Significant gender differences in motivation to lead were found in areas related to self-efficacy, prior leadership experience, and personality traits of extraversion and individualism. There were no significant gender differences in motivation to lead related to personality traits of collectivism, agreeableness, and emotional stability.