Cryopreservation of HEP-G2 Cells Attached to Substrates: The Benefit of Sucrose and Trehalose in Combination with Dimethyl Sulfoxide

Abstract

BACKGROUND: Cryopreservation of mammalian cells is mainly done in cryovials as free cell suspension in 5 to10% (v/v) dimethyl sulfoxide (DMSO). Relatively little attention has been paid to cryopreservation of adherent cell monolayers. OBJECTIVE: To investigate the appropriate cryoprotectant (CPA) formulations for the cryopreservation on HEP-G2 human tumor cells attached to the polystyrene plate and plastic surfaces. MATERIALS AND METHODS: Five CPA formulations were evaluated for the cryopreservation of HEP-G2 cells attached to polystyrene plates and and plastic coverslips, using post-thaw cell viability as a performance indicator. RESULTS AND CONCLUSION: Hep-G2 cells attached to the plastic coverslips and polystyrene plate surfaces were successfully cryopreserved in 10% DMSO with sucrose and trehalose. The addition of saccharides enabled the reduction of DMSO concentration, replaced serum, and improved the functional capacity of post-thaw Hep-G2 cells. Cells attached to the plastic coverslips show significantly better results than those attached to the polystyrene plate surfaces after cryopreservation.