In vitro study on cryopreservation of peripheral blood stem cells with uncontrolled freezer

Abstract

Objective:To evaluate the effects of cryopreservation of APBSCs in -80"C un-controlled freezer and liquid nitrogen, and to search for the efficient combined cryoprotectant and the highest cell concentration for cryopreservating peripheral blood stem cell (PBSC). Methods: To compare the effect of three combined cryoprotectants, evaluation of in vitro proliferative capacity by colony-forming unitGranulocyte-macrophage (CFU-GM) and burst-forming Unit-erythroid (BFU-E) assay, immunophenotyping for CD34+/CD38 cells by FCM, and viability assessment by trypan blue exclusion were performed. Results: The cryoprotectant 10%DMSO+HES+auto-Plasma resulted in the highest recovery rates. CFU-GM and BFU-E were (78.7+9.8)%, (69.8+14.1)%, respectively. The recovery rates of CFU-GM and BFU-E in A/C groups were (68.3+6.2)%/ (65.8:t=7.2)% and (63.4+9.7)%/ (60.4+ 10.5)%, respectively. The recovery rates of CD34+/CD38 cells and cell viability with the three combined reagents were 90% and 80%, respectively. Conclusion: 5%DMSO+ HES+auto-PLASMA is an ideal combined cryoprotectant for cryopreserving PBSC. The cell concentrations may be cryopreserved at 4xl08 ml l.

Guidelines to Optimize Survival and Migration Capacities of Equine Mesenchymal Stem Cells

“…In addition, temperature-controlled thawing did not influence the cell viability in comparison to hand-thawing. In this regard, it has been described that thawing human peripheral blood stem cells (PBSCs) at 20°C or 37°C did not influence cell recovery or viability [17,18]. Therefore, we suggest that hand thawing is a suitable method to be used in the field.…”