51 species of lignicolous macromycetes were isolated from natural substrates, screened and then were preserved through in vitro culturing and cryopreservation. The isolation was performed using dikaryotic mycelium from the fruit-bodies and the pure cultures were deposited in the Culture Collection of Research Laboratory for fungi with applications in ecological reconstruction of heavy metals polluted soils – RECOSOL. In this paper, the in vitro characterization of 8 isolates is presented. The mycelium’s growth rhythm and macroscopic features of the colony, along with microscopic features of mycelium were recorded. The cryopreservation of fungal mycelium was performed using glycerol 10% as cryo-protective agent, after the cultivation of mycelium onto liquid media and homogenization. Cryopreservation proved to be effective.