UM171和SR1对不同来源造血干/祖细胞体外扩增的作用研究

目的探讨小分子化合物UM171和SR1对脐带血、供者动员外周血和淋巴瘤患者自体动员外周血3种来源的造血干/祖细胞(HSPCs)体外扩增的作用。方法将3种来源的CD34+细胞分别予以UM171、SR1干预后进行体外扩增培养,记为对照组、UM171组、SR1组和UM171+SR1组。通过细胞计数检测各组总有核细胞的数量,流式细胞术检测HSPCs的比例、各谱系分化细胞的比例和HSPCs上归巢相关因子CXCR4的表达水平。多组数据若满足方差齐性,采用单因素方差分析,组间两两比较采用LSD-t检验;若方差不齐,多组间比较以及两两比较均采用Kruskal-Wallis检验。结果与对照组比较,UM171和SR1均能促进3种来源HSPCs的比例升高,同时UM171能够增加3种来源HSPCs的扩增倍数。与对照组比较,UM171处理后脐带血来源的CD33^+(髓系)细胞的比例升高,CD41^+(巨核)细胞的比例降低;SR1处理后3种来源的CD3-CD56^+(自然杀伤)细胞的比例均升高。体外扩增48 h后各组HSPCs上CXCR4的表达较培养前增加。结论UM171能够有效扩增3种来源HSPCs的数量,促进脐带血来源HSPCs分化为髓系细胞并抑制其分化为巨核细胞。SR1能够促进3种来源HSPCs分化为自然杀伤细胞。体外扩增培养可以提高3种来源HSPCs上CXCR4的表达水平。

Effects of UM171 and SR1 on ex vivo expansion of hematopoietic stem and progenitor cells from different sources

ObjectiveTo investigate the effects of small molecule compounds UM171 and SR1 on hematopoietic stem and progenitor cells (HSPCs) from umbilical cord blood (CB) , donor mobilized peripheral blood (Donor-mPB) and autologous mobilized peripheral blood (Auto-mPB) . MethodsHSPCs from three sources were cultured with or without UM171 and SR1, and named control group, UM171 group, SR1 group and UM171+SR1 group.The number of total nucleated cells in each group was detected by cell counting. The proportion and differentiation of HSPCs and the expression of CXCR4 on HSPCs were detected by flow cytometry.One-way ANOVA and Kruskal- Wallis test were used for statistical analysis. ResultsCompared with control group, the proportions of HSPCs from three sources were increased both after treatment of UM171 and SR1. And the treatment of UM171 could increase the expansion folds of HSPCs from three sources significantly. Compared with control group, UM171 treatment led an increase in the proportion of CD33⁺ (myeloid) cells as well as a decrease in the proportion of CD41⁺ (megakaryocyte) cells from CB-derived HSPCs, SR1 treatment led an increase in the proportion of CD3^- CD56⁺ (natural killer) cells from HSPCs from three sources. The expression of CXCR4 on HSPCs in each group increased significantly after 48 h of ex vivo expansion relative to that before culture. ConclusionUM171 could effectively expand the number of HSPCs from CB, Donor-mPB, Auto-mPB and promote the differentiation of CB-derived HSPCs into myeloid cells as well as inhibit their differentiation into megakaryocytes. SR1 could promote the differentiation of HSPCs from three sources into natural killer cells. The expression of CXCR4 on HSPCs from three sources was increased significantly after ex vivo expansion.