[Measures for Maintaining the Vitality of Peripheral Blood Hematopoietic Stem Cells in vitro].

OBJECTIVE: To explore the maintaining measures for the vitality of hematopoietic stem cells (HSC) in vitro, so as provide technical support for ultra long distance transport of HSC collected from unrelated donors.

METHODS: Peripheral blood hematopoietic stem cells (PBHSC) were treated by different methods according to various groups, then stored at 4 ℃ in the refrigerator. The percentage of CD34+ cells, relative cell activity, relative cell proliferation rate, relative colony-forming rate, oxygen fraction and intracellular reactive oxygen species (ROS) were detected at 0, 24, 48 and 72 h after storage of PBHSC respectively.

RESULTS: The percentage of CD34+ cells during 72 h storage did not altered. Along with the prolonging of storage time, the relative cell activity, relative cell proliferation rate and relative colony-forming rate gradually decreased in untreated PBHSC(control group), the related coefficients were -0.796, -0.883 and -0.815 respectively. Plasma dilution, antioxidants and oxygenation could improve the relative cell activity and relative cell proliferation rate, but oxygenation could decrease the relative colony-forming rate of PBHSC. The combination of 2 or 3 factors showed stronger protection effects on PBHSC. The intracellular level of ROS decreased gradually with the prolonging of storage time. Oxygenation of PBHSC could increase oxygen fraction, and also increase the intracellular level of ROS at the same time. The addition of antioxidants could reduce the level of ROS.

CONCLUSION: The percentage of CD34+ cells can not serve as the indicator of PBHSC vitality. Plasma dilution, oxygenation and antioxidants can increase the survival and viability of PBHSC, but oxygenation can increase the intracellular ROS level and impair colony-forming ability of PBHSC. The combination of multiple factors can maintain the vitality of PBHSC better.