Comprehensive study of hydrostatic pressure treated human umbilical cord blood cells via response surface method

Abstract

Amelioration of the survival parameters of cryopreserved samples after thawing has already been addressed through several techniques including vitrification to avoid the formation of ice cores. However, this approach cannot be followed in the case of samples with higher volumes. Hydrostatic pressure (HP) treatment has been proven to increase some qualifying parameters (e.g., motility, insemination efficiency) of certain biological samples. Accordingly, the preparation of umbilical cord blood (UCB) samples through an active (mechanical) pre-stressing process to increase the survival rate of cryopreserved samples can be regarded as a novel strategy that calls for basic experimental studies. The goal of our study was to assess the effects of HP treatment on the qualifying parameters (DNA fragmentation by agarose gel electrophoresis and capillary electrophoresis, Total Nucleotide Cell (TNC) count, CD34+/CD45+ count, and superoxide dismutase activity (SOD) of human umbilical cord blood (UCB) derived cells). The experimental arrangement was set to provide data for response-surface analysis to take into account the common effects of the individual variables of pressure and time exposure. 3D visualization of experimental data revealed that 50-min long HP treatment at 12.5MPa can significantly (α=0.05) enhance white blood cell (WBC) and CD34+/CD45+ cell counts. However no DNA fragmentation was observed even at higher pressures, SOD activity was triggered over 15.0MPa. As a conclusion, HP treatment may contribute to the optimal cryopreservation of UCB cells by significantly increasing WBC and CD34+/CD45+ cell counts without adverse effects neither on DNA stability nor on triggering SOD activity.