Dual dependence of cryobiogical properties of Sf21 cell membrane on the temperature and the concentration of the cryoprotectant.

Abstract

The Sf21 cell line is extensively used for virus research and producing heterologous recombinant proteins. To develop optimal strategies for minimizing cell injury due to intracellular ice formation and excessive volume shrinkage during cryopreservation, the fundamental transport properties including the osmotic inactive volume (Vb ), the hydraulic conductivity (Lp ), and the glycerol permeability (Ps ) of Sf21 cell membrane at 25, 15, 5 and -2°C were characterized using a micro-perfusion chamber. The effects of temperature on the hydraulic conductivity and the glycerol permeability of Sf21 cell membrane, reflected by the activation energies, were quantitatively investigated. It was found that the hydraulic conductivity decreases along with the increase of the final CPA concentration at a given temperature, and quantitative analysis indicates that the hydraulic conductivity has a significant linear attenuation along with the increase of the concentration of glycerol. Therefore, we incorporate the concentration dependence of the hydraulic conductivity into the classic Arrhenius relationship by replacing the constant reference value of the hydraulic conductivity at the reference temperature with a function that is linearly dependent on the CPA concentration. Consequently, the prediction of the Arrhenius relationship is improved, and the novel Arrhenius relationship could be very important to the development of optimal strategies for cell cryopreservation.