The Influence of Different Concentrations of Cryoprotectants on Neuronal Cell Viability

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.