Aim: The purpose of this study was to compare clinical and laboratory characteristics of infertile males according to their postwash progressively motile sperm count and to evaluate whether there was a relationship between serum vitamin D (VD) levels and semen parameters. Material and Method: A total of 198 infertile men were included in this cross-sectional study. Study population was mainly divided into two groups according to post wash total progressively motile sperm count (TPMSC) as less than 5 million/ml (study group) and equal or greater than 5 million/ml (control group). The main parameters recorded for each patient were; age, BMI (body mass index), infertility type, infertility duration, previous operation, history of disease, smoking, drug usage and serum levels of 25OHVD3, total calcium (Ca) and testosterone (TT), gonadotropins, and semen parameters. Results: There were no statistically significant differences between groups in terms of age, BMI, infertility type, infertility duration, and previous operation, history of disease, smoking, drug usage, TT, and 25OHVD3 levels. Serum levels of gonadotropins were significantly lower in the study group (p<0.001). The mean 25OHVD3 level was 21.0 +/- 7.2 ng/ml in the total group and there was a positive correlation between VD and pre/post wash sperm count in the study group (p=0.036, p=0.034, respectively). Serum Ca level was also significantly lower in this group (p=0.012). Discussion: Vitamin D insufficiency was common among the infertile men. VD and Ca supplementation may be appropriate treatment for infertile males with documented VD and Ca deficiency, whose post wash TPMSC lesser than 5 million/mL. Further well designed and large series prospective controlled studies are needed on this issue.