Cryopreservation of human whole blood and checking the viability of mononuclear cells

Abstract

Now a days, cryopreservation of whole blood & whole blood derived mononuclear cells (MNC’s) has emerged as a promising technique for the healing of the subjects going through various blood related ailments. The supremacy of this technology are amazing cell viability, the cell traits remain intact i.e. the retention of the structural and functional intactness of the super freezed cells after decades of preservation However, there are a host of challenges in cryopreservation while preserving the cells due to different levels of attached intricacies that involves achieving cell viability, cell function, cell proliferation and differentiation. The present work investigated the outputs using some selected cryoprotectant on post thawing viability mononuclear cells (MNCs) derived from peripheral whole blood during cryopreservation process along with just preserving peripheral whole blood. The extracellular cryoprotectant such as Dextran and HES with a concentration of 10% w/v and intracellular cryoprotectant such as glycerol and DMSO with a concentration of 10% v/v were used. Post thaw cultured samples were analyzed for cell viability by trypan blue membrane exclusion assay. Samples consisting of hydroxy ethyl starch (HES) were found to be the most effective cryoprotectant achieving average cell viability of 84 %. Whereas only 63% cell viability was obtained using Dextran as cryoprotectant. SEM and phase contrast microscopy confirm the normal cell morphology of the post thaw cultured whole blood and mononuclear cells respectively