Isolation and propagation of the animal rotaviruses in MA-104 cells - 30 years of practical experience.

•Based on the experiences of 30 years we describe an optimal protocol for isolation and propagation of rotaviruses (RVs).•MA-104 cells were successfully for the isolation and propagation of RV strains of species A (RVA).•The centrifugation of prepared specimens (faeces, gut contents) on the cells promotes the virus entry (adsorption and penetration).•The combination of the activation of virus particles with pancreatic enzymes (trypsin) prior to their inoculation on cells, the pre-treatment of cells with diethylaminoethyl (DEAE)-dextran solution, and the use of roller culture technique is the method of choice.•All 102 cell culture-adapted virus strains belonging to RV species A, while non-RVA strains could not be isolated.