枸杞多糖联合人骨骼肌源性血管外膜细胞对人脐血CD34+细胞造血支持的体外研究  

Effect of Lycium Barbarum Polysaccharide Combined with Human Skeletal Muscle-derived Pericytes/Perivascular Cells on Hematopoietic Support of Umbilical Cord Blood CD34+ Cells in Vitro

作　　者：张磊[1] 张思齐 杨婷婷[2] 马洁[3] 郑波 ZHANG Lei;ZHANG Siqi;YANG Tingting;MA Jie;ZHENG Bo(Ningxia Medical University,Yinchuan 750004,China;Human Stem Cell Institute,the General Hospital of Ningxia Medical University,Yinchuan 750004,China;Department of Hematology,the General Hospital of Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学,银川750004 [2]宁夏医科大学总医院人类干细胞研究所,银川750004 [3]宁夏医科大学总医院血液内科,银川750004

出　　处：《宁夏医科大学学报》2022年第6期547-554,共8页Journal of Ningxia Medical University

基　　金：国家自然科学基金项目(8186010202);宁夏自然科学基金项目(2020AAC03370)。

摘　　要：目的探讨枸杞多糖(LBP)对人脐血CD34^(+)造血干/祖细胞(UCB CD34^(+)HSPCs)体外扩增的效力。方法利用CCK-8法检测不同浓度LBP对CD146^(+)hMD-PCs增殖率的影响;流式细胞术检测LBP对CD146^(+)hMD-PCs细胞周期的影响;UCB CD34^(+)HSPCs的体外共培养实验分为LBP^(+)CD146^(+)hMD-PCs组、LBP组、CD146^(+)hMD-PCs组和空白对照组,于培养的1、2、4周分别对各组扩增后细胞数量、集落数量和免疫表型进行分析比较。结果CCK-8及流式细胞周期检测显示,与未添加LBP组比较,当LBP浓度为150 mg·L^(-1)时,CD146^(+)hMD-PCs细胞活力升高,G0/G1期细胞比例减少,S期、G2/M期细胞比例升高(P均<0.01)。共培养第1、2、4周后,LBP^(+)CD146^(+)hMD-PCs组细胞数量均多于CD146^(+)hMD-PCs组及LBP组,CD45^(+)、CD34^(+)CD33^(-)、CD10^(+)CD19^(+)、CD14^(+)细胞比例均高于LBP组,且LBP^(+)CD146^(+)hMD-PCs组集落形成单位数量均多于CD146^(+)hMD-PCs组(P均<0.05);与CD146^(+)hMD-PCs组相比,LBP组在共培养第1、2、4周后细胞数量均降低,CD45^(+)、CD34^(+)CD33^(-)、CD14^(+)细胞比例均降低;CD10^(+)CD19^(+)细胞比例在共培养第2、4周后均降低(P均<0.05);在共培养第2周时,LBP组细胞数量多于空白对照组(P<0.01)。结论LBP可以通过促进CD146^(+)hMD-PCs的增殖从而促进UCB CD34^(+)HSPCs的体外扩增。Objective To investigate the efficacy of Lycium barbarum polysaccharide(LBP)on the expansion of human umbilical cord blood CD34^(+)hematopoietic stem/progenitor cells(UCB CD34^(+)HSPCs)in vitro.Methods CCK-8 method was used to detect the effect of different concentrations of LBP on the proliferation rate of CD146^(+)hMD-PCs.Flow cytometry was used to detect the effect of LBP on the cell cycle of CD146^(+)hMD-PCs.The co-culture experiment of UCB CD34^(+)HSPCs in vitro was divided into 4 groups:LBP^(+)CD146^(+)hMD-PCs group,LBP group,CD146^(+)hMD-PCs group,and blank control group.The number of expanded cells,colonies and immunophenotypes were analyzed and compared at 1,2 and 4 weeks.Results CCK-8 and flow cytometry showed that compared with the group without LBP,when the concentration of LBP was 150 mg·L^(-1),the cell viability of CD146^(+)hMD-PCs was increased significantly;the proportion of G0/G1 phase cells decreased,and the ratio of S phase and G2/M phase cells increased(P all<0.01).The co-culture results of LBP^(+)CD146^(+)hMD-PCs group comparing with CD146^(+)hMD-PCs group and LBP group,showed that the number of cells increased significantly;the expression levels of CD45^(+),CD34^(+)CD33^(-),CD10^(+)CD19^(+)and CD14^(+)cells increased in varying degrees of co-culture after 1,2 and 4 weeks;the number of colony forming units in LBP^(+)CD146^(+)hMD-PCs group was higher than that in CD146^(+)hMD-PCs group(P all<0.05).The co-culture results of LBP group comparing with the CD146^(+)hMD-PCs group,demonstrated that the number of cells and the proportions of CD45^(+),CD34^(+)CD33^(-),and CD14^(+)cells in the LBP group decreased significantly after the 1,2,and 4 weeks of co-culture,and the proportion of CD10^(+)CD19^(+)cells decreased after 2 and 4 weeks of co-culture(P all<0.05).The co-culture results of LBP group comparing with blank control group,indicated that the number of cells increased after co-culture for 2 weeks(P<0.01).Conclusion LBP can promote the expansion of UCB CD34^(+)HSPCs in vitro by promoting the p

关 键 词：枸杞多糖 人骨骼肌源性血管外膜细胞 造血干/祖细胞 造血支持 

分 类 号：R285.5[医药卫生—中药学] R551[医药卫生—中医学]