Aim. To characterize a new hormone-independent strain.-27. of R. serpentina tissue culture. Methods. Plant tissue culture, biochemical and cytological analysis, statistical methods. Results. The strain is maintained on the specially designed simple-composition hormone-free 10C medium. The maximum biomass yield was at day 69 of subculture (693.5 g/l of live biomass, 43.0 g/l of dry biomass), the weight of cell biomass increased 15-18 times over a subculture period. The maximum content of indole alkaloids was observed from 88 to 108 days of sub-culture. The dry biomass contained 4.0% of total indole alkaloids, 1.64% of ajmaline-like alkaloids, 0.789% of ajmaline, 0.337% of vomilenine, and 0.006% of each yohimbine and reserpine. The strain is a heterogeneous mixoploid cell population with a modal class of 22 to 33 chromosomes (2n = 22), which characterized by significant cell and nuclear polymorphism. Conclusions. The K-27M strain of R. serpentina cultured tissue is a prospective producer of indole alkaloids.