Role of N-acetyl cysteine on bone marrow-derived hematopoietic stem/progenitor cell cryopreservation: Outcome on the oxidative stress-mediated cryodamage and repopulation Capacity into hematopoietic lineages

ABSTRACT

Cryopreservation of hematopoietic stem/progenitor cells (HSPCs) is associated with oxidative stress-mediatedcryodamage, hence compromising the therapeutic potency. The roles of N-acetyl cysteine (NAC) on the oxidativestress-mediated cryodamage and repopulation capacity of HSPCs into myeloid, erythroid, and pre-B lymphoidprogenitors were investigated. Mice bone marrow-derived HSPCs were cultured for 24 hours, followed bycryopreservation at 1 × 106/ml cells in cryomedium containing 10% dimethyl sulfoxide with NAC (0.25, 0.5, or 2.0µM) or without for 48 hours, 2 weeks, and 4 weeks at −80°C. Cryopreservation significantly reduced cell viability atpost-thawed (p < 0.05) with long-term cryopreservation conferred a greater cell recovery. NAC improved the bonemarrow-derived HSPC viability (p < 0.05) at 0.5 and 2.0 µM after 48 hours of cryopreservation. Cryopreservationlowered the malondialdehyde level (p < 0.05) although glutathione, superoxide dismutase, and protein carbonyl levelswere not significantly affected. Repopulation capacity of HSPCs into myeloid–erythroid progenitors was greatlyreduced (p < 0.05) after 4 weeks of cryopreservation as compared to precryopreservation group. Meanwhile, NACsupplementation showed no remarkable effect on oxidative stress-mediated cryodamage and repopulation capacity ofHSPCs. Conclusively, the cryoprotective role of NAC on the cryopreservation of HSPCs deserves further investigation