Seven wheat genotypes were used in this investigation to study the effect of genotypes, levels of polyethylene glycol (PEG) concentrations and their interaction on immature embryo induction, as well as, estimating heritability for in vitro traits. The genotypes were in vitro evaluated for immature embryos culture ability at four levels of PEG concentrations. Significant differences between genotypes were obtained for some traits at four PEG levels. Levels of mean squares were significant for some traits such as number of green point, plant length and number of tillers. Whereas, mean squares of interaction between genotypes × PEG levels was insignificant for all traits except number of tillers. The results demonstrated that the greatest percentages for green points and plant weight were recorded by the hybrid Sakha 93 × Gemmiza 9. In addition, Sakha 93 × Sids 1 was the best genotype for root length and number of tillers. Whereas, the greatest value for number of plantlets was observed in Sakha 93 and sakha 94. The addition of 10 g/l PEG to MS nutrient medium and 1 mg/l NAA and 0.5 mg/l Kinetin in regeneration medium could be the best concentration for immature embryo purpose in wheat. Furthermore, the genetic variations were highly positive for some traits such as number of green point at levels A and B, number of plants at levels A,B and D, plant weight at level A, plant length at level C, root length at level A and number of tillers at levels A,B and D. The values of heritability were ranged between 53.66 to 90.48% for number of tillers at levels D and A, respectively. The results indicated that the genes controlled growing traits in wheat were highly affected by media composition.