An accurate and efficient method for measuring the mass of hairy roots using conductometry is established. A conductivity equation expressed in terms of the concentration of the ion species in the medium is suggested. By using this equation, the effect of the individual ions on the total conductivity can be quantitatively analyzed. An equation for the in situ estimation of the cell growth coefficient for determining the mass of hairy roots is established based on measurements of the nitrogen concentration and conductivity during cultivation. The proposed equation does not require preliminary experiments to determine the cell growth coefficient. Instead, the physiological characteristics of the plant species are reflected by introducing the cellular nitrogen content. Since the cell growth coefficient is determined by measuring the major ionic nutrient concentrations, it is more effective to express the dynamics of an actual culture system. This improved method for determining the mass of hairy roots was successfully utilized in a fed-batch culture system.