The Influence of Different Concentrations of Cryoprotectants on Neuronal Cell Viability

Kim, Young Soo; Han, Ki Soo; Lee, Uhn; Kim, Young Bo; Yoo, Young Mi

Abstract

Apromising technique for the treatment of Parkinson's disease and other various neurodegenerative disorders is the transplantation of fetal neural tissue. There must, however, be a prompt and reliable source, and one solution is cryopreservation, where tissue viability can maintained for prolonged periods. Fetal neural tissue is, however, known to be susceptible to freeze-storage damage during cryopreservation. In this study, we examined the influence of different concentrations of cryoprotectants upon the survival of rat fetal neurones. Fetal rat brain tissue was frozen with 7-15% dimethyl sulfoxide(DMSO) and 10-50% fetal bovine serum(FBS) as cryoprotectants, then stored for a period of 5 months. Post-storage neuronal cell viabilty was assessed by vital staining followed by determination of cell density. Average total viability of frozen cells with 7% DMSO and 10-50% FBS was less than 50%. Cryopreserved cells with 10-50% DMSO and 10-50% FBS showed almost the same viability(around 70%). The highest viability was obtained with 15% DMSO+20% FBS combination(76%) and 10% DMSO+10% FBS combination(75%). Theoretically, the higher the concentration of cryoprotectants, the higher the viability: however, the best result was achieved stated above, when the combination of cryoprotectants was at the concentrations stated above.