Mammalian sex hormones (MSHs) are synthesized by various plant species and known play critical roles in growth, development, and reproduction. External and endogenous use of MSHs resulted in different response in different plant species. However, effects of different MSHs with concentrations on different wheat genetic backgrounds have not been studied in vitro. Therefore, three wheat cultivars (cvs. Palandöken 97, Nenehatun, Kırik) and four MSHs (17 β-estradiol, estrogen, progesterone, and testosterone) with three concentrations of 10-4, 10-5, 10-6 mM were used to investigate callus formation and embryonic callus formation rates as well as number of regenerated plants. Our results showed that wheat genotypes were significantly influenced by the type of MSH and its concentration. In addition, effect of hormones may vary with respect to growth or development of cells or plants. The highest callus formation rate was obtained on cv. Palandöken 97 with 10-4 mM testosterone hormone. Embryogenic callus formation rate with respect to number of explants was the highest on Palandöken 97 in 17β-estradiol with 10 -4 mM while embryogenic callus formation rate with respect to number of calli was the highest on cv. Kırik in progesterone and 10-4 mM concentration. On the other hand, the highest regenerated plants were observed on cv. Palandöken 97 in 10-6 mM 17β-estradiol and 10-6 Mm progesterone. Such contrasting results may indicate the potential utilization of suitable wheat genotypes, MSHs and doses in wheat tissue culture which is a prerequisite in wheat transformation experiments.