The aim of the chapter is to compare the level of employee motivation in enterprises in Slovak and the Czech Republic. Sampling unit consists of 4444 respondents from Slovakia and 2312 from the Czech Republic. Following our outcomes, we can state that the most important motivation factors are mostly identical in both countries; however, there is a slight difference in the order of their importance. Motivation factors relating to financial reward are the most important for employees in the Czech Republic. Basic salary is a motivation factor important mainly for women in Slovakia. Demands of Czech respondents are higher in motivation factors relating to interpersonal relationships. In general, we can state that, in terms of gender, needs of women are more exacting than those of men. Dependence between two categorical variables was verified using Pearson's chisquare statistics. We found out that despite big similarities in the order of importance of individual motivation factors, there are significant differences between selected motivational needs of employees in individual countries.