Paphiopedilum concolor (Bateman) Pfitzer is a native slipper orchid of Northern Vietnam. Currently, this species has been almost extinct in the wild due to deforestation and overexploitation, and therefore, it is very important to develop an efficient in vitro culture method for conservation. This paper reports the factors affecting high frequency germination and regeneration in vitro of Paphiopedilum concolor, including seed maturity, basal nutrient medium, plant growth regulators and additive organic compounds. The seeds at the age of 190 days after pollination (DAP) showed the highest germination rate (85.0%) and the germination time was 40 days. The germination rates on both half-strength MS and half-strength VW media reached 73.8% and 72.5%, respectively. The optimal medium for protocorm-derived shoot regeneration and growth of planlets was half-strength MS medium supplemented with 1.0 mg l^-1 BAP, 0.5 mg l^-1 NAA, 100 ml l^-1 coconut water, 40 g l^-1 potato homogenate, and 30 g l^-1 banana homogenate. On this medium the average number of shoots and leaves was 3.3 and 3.6, respectively. The leaves were thick, large, and dark green. Their length reached 1.5 cm. The best rooting effect was observed on the half-strength VW medium supplemented with 1.0 mg l^-1 NAA combined with 0.5 mg l^-1 AC, and 30 g l^-1 banana homogenate. The average number of roots reached 4.5, increasing 2.5 roots compared to the control.