Dendrobium wardianum R. Warner is arare orchid species in Thailand. Efficient propagation method is essential for the conservation of this orchid. Inthis study, effects of plant growth regulators (PGRs), i.e., 6-benzylaminopurine (BA) and α-naphthalene acetic acid (NAA), and organic substances, i.e.,tryptone, yeast extract and/or coconut water, on seed germination and seedling development of D. wardianum were investigated. The germination rate of D. wardianum seeds at two months after sowing was either insignificantly affected or slightly hindered by the addition of the studied PGRs or organic substances. However, these supplements couldgreatly improve protocorm and further seedling development. Theaddition of 2 g/l of either tryptone or yeast extract to Murashige and Skoog medium (MS) resultedin the highest numbers of green protocorms (i.e., 43.46% and 38.65%, respectively) and protocorms with shoots and rhizoids (i.e., 1.97% and 3.44%,respectively) compared to 11.13% of green protocorms and 0% of protocorms with shoots and rhizoids on MS without addition. Moreover, half-strength MS (½MS) supplemented with either 2 or 4 mg/l of BA, 2 g/lof yeast extract and 2 g/l of tryptone or ½MS supplemented with 150 ml/l of coconut water could significantly inducethe number of shoots per plantlet (2.90–3.73 shoots) comparedto ½MS (1.14 shoots). Therefore,this study showed that the addition of PGRs and/or organic supplements to the basal medium could improve asymbiotic seed germination of D. wardianum.