Due to the increasing demand and over collection of orchids from nature to produce salep, scientists have been led to search for more efficient ways to propagate these specific orchids in vitro. This present study compares germination performances of two commercial (Orchimax and Knudson C) and one specially prepared orchid growth media (SV), on economically and medicinally important orchids used to make salep; (Anacamptis pyramidalis, Dactylorhiza romana, Neotinea tridentata and Anacamptis morio subsp. morio) and further aims to obtain a mature orchid plant by following the natural environmental cycle. Significant differences in seed germination and protocorm development were observed. Asymbiotic germination tests showed that the specially prepared growth media performed better than the commercial media by 79.11% germination rate. Also, that A. morio subsp. morio had the best germination rate by 88.91%. Protocorms developed in the sixteenth week after sowing. Soil was collected from the natural habitat of each species and was used as a potting substrate, and this helped orchids to pass their initial acclimatization stage. Regeneration success of orchids at in vitro conditions could be increased by using SV growth medium, following their natural hardening off cycle and using specific substrates from their respective habitats.