Paphiopedilum is an ornamental orchid used mainly for interior decoration. As the division of plants is uneconomical, a fast method of propagation is needed. The aim of this study was to evaluate the influence of medium type (MS or VW), growth regulators, i.e. BA, KIN, TDZ used separately or in combinations and natural additives, i.e. coconut water, banana pulp, casein hydrolysate, on Paphiopedilum insigne plantlets grown in vitro. It was found out that BA in concentration of 0.5 mg·dm–3 allowed to obtain the highest multiplication rate (2.92), however the use of KIN in concentration of 1 mg·dm–3 resulted in formation of bigger and higher quality plantlets, It is possible to replace cytokinins with other biologically active substance, such as banana pulp. The 1/2 MS medium might be used for Paphiopedilum insigne tissue culture, as there was no difference in terms of multiplication rate and the obtained plantlets were of better quality, especially that it is cheaper and easier to prepare.