The research, entitled "The Use of Organic Fertilizer and Cherry Extract (Muntingia calabura L.) for Medium Substitution VW (Vacint And Went) and Sucrose in Subculture Medium Vanda tricolor Orchids In Vitro" was studying the effect of organic fertilizers and cherry extract, and determining the concentration of organic fertilizer and cherry extract that will be an effective formula to substitute medium subculture of Vanda tricolor. The research was conducted in January to April 2016 at Laboratorium of Vitro Culture, Faculty of Agriculture, Universitas Muhammadiyah Yogyakarta. This research was design by using Completely Randomized Design (CRD) with 10 replications and was done by using a single factor experimental design.The treatments were organic fertilizer 3 ml / L and cherry extract with variety of 50g / L, 100g / L, 150g / L, and 200g / L. The parameter of observation were the height of shoots, increasing number of leaves, increasing number of shoots, increasing number of shoots apical, increasing number of roots, percentages of live explants, percentages of explants contamination and percentages of explants browning. The results revealed that the use of organic fertilizer 3ml/L+ sucrose 30g/L could replace Medium VW+ sucrose 30g/L and revealed the best result in subculture of Vanda tricolor