In a world where population is outgrowing land availability for food supply, plant biotechnology needs to be addressed and implemented as an aid to higherproductivity. The continuous use of antibiotics has also resulted inmultiresistant bacterial strains all over the world.As a consequence, growing research has taken place to look for naturally produced plant compounds with antibiotic properties. In the last years, new natural peptides, among them the highly interesting group of cyclotides, have thus been discovered. They have shown antimicrobial properties while posing no harm to mammalian cells.As of today, a systematic analysis of significant variables for an optimal plant cell culture growth and differentiation aiming at high productivity has not been fullyestablished yet.This study intends to partially solve this problem: key variables of the process are correlated to obtain a matrix ofdifferentiated cells of Viola tricolor, finally proceeding with the setup of organ suspension cultures under optimized conditions to yield highly useful cyclotides.