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Effect of Simulated Microgravity on the Differentiation of Hematopoietic Stem and Progenitor Cell into Functionally Mature Neutrophil
Wang Zhezhe, Zhang Xiaoyu, Zhang Sudong, Xie Xinyan, Liu Peng, Xiang Fei, Ren Qian, Xie Xuemei, Guo Rongxia, Liu Fei, Wang Tong, Xu Yuanfu*
State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China
Abstract: This paper aims to determinate the impact of the RCCS (rotate cell culture system) on granulopoiesis of hematopoietic stem/progenitor cells and evaluate the efficiency of functionally mature neutrophil generation. CD34+ cells enriched from umbilical cord blood were cultured in StemSpan SFEM containing SCF (stem cell factor)， Flt3 (Fms-related tyrosine kinase 3 ligand)， IL-3 (interleukin-3) and TPO (thrombopoietin). After 7 days， cells were cultured in medium containing SCF， Flt3， IL-3， IL-6 and G-CSF (granulocyte colony stimulating factor). After 7 days of expansion， those cells were divided into 2 groups， SC (static culture) group and RCCS group， and separately cultured for next 9 days inducing granulocyte differentiation. To evaluate the differentiate efficiency， the expression of surface markers， such as CD34， CD38， CD11b， CD16b and CD66b were detected by flow cytometry. On day 16， cells were harvested and numerical expansion， cell morphology， chemotaxis， phagocytosis and ROS production were used for evaluating the quality and quantity of neutrophils from the two groups. Both groups could successfully induce functionally mature neutrophils， while the percentage of CD16b+ cells in RCCS group was higher than that in SC group (29.82%±2.48% vs 15.01±0.62%， P<0.01). Compared to the SC group， RCCS could increase the efficiency of neutrophil ROS production and chemotaxis (P<0.05). Altogether， these results indicated that the RCCS significantly promoted the efficiency of generation of functionally mature neutrophil from hematopoietic stem and progenitor cells.
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