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作 者:区文超[1] 鲁树坤[1] 张芳婷[2] 张立兵[2] 聂李平[3] 周小莹[4] 谭孟群[4]
机构地区:[1]深圳北京大学香港科技大学博士后工作站、北京大学深圳医院超声影像科,广东深圳518036 [2]深圳北京大学香港科技大学博士后工作站、北京大学深圳医院中心实验室,广东深圳518036 [3]深圳北京大学香港科技大学博士后工作站、北京大学深圳医院检验科,广东深圳518036 [4]中南大学湘雅基础医学院生理系血液生理研究室,湖南长沙410078
出 处:《中国病理生理杂志》2008年第2期357-360,共4页Chinese Journal of Pathophysiology
基 金:广东省自然科学基金资助项目(No.31005);深圳市科技计划项目重点课题资助项目(No.JH200505270412B)
摘 要:目的:探讨干细胞因子(SCF)+白细胞介素-6(IL-6)短期扩增对CD34+造血干/祖细胞黏附和迁移能力的影响。方法:用密度剃度离心的方法分离脐血CD34+细胞,经SCF和IL-6孵育48 h,用CCK-8方法检测CD34+细胞增殖能力;用流式细胞仪检测处理前后的CD49d(VLA-4)、CD11 a(LFA-1)、CD62L(L-selectin)及CD184(CXCR4)的表达。用纤连蛋白(FN)包被96孔板,检测经或未经因子扩增的CD34+细胞的黏附能力。扩增的CD34+细胞悬浮于transwell培养板的上层,下层添加基质细胞衍生因子(SDF-1),流式细胞仪检测迁移细胞数,计算迁移率。结果:经SCF+IL-6处理48h后CD34+细胞扩增近3倍;表达CD49d、CD11 a、CD62L及CD184的CD34+细胞的百分数分别由原来的26.34%±5.37%、17.63%±4.57%、46.38%±6.61%和9.58%±1.56%增加到65.67%±8.72%、56.67%±6.34%、84.76%±9.57%和19.32%±3.64%(P<0.01)。扩增后的CD34+细胞对FN的黏附能力及在SDF-1诱导下的迁移作用都显著增强(P<0.01)。结论:SCF+IL-6短期扩增CD34+造血干/祖细胞显著增加细胞的黏附能力,增加SDF-1诱导的迁移作用,可能是SCF+IL-6促进归巢的主要机制之一。AIM: To investigate the expression and function of homing related molecules and transmigration a- bility of human cord blood CD34^+ hematopoietic stem/progenitor cells after short time stimulation with cytokine SCF and IL - 6. METHODS: CD34^+ cells were separated by Ficoll density gradient centrifugation and stimulated by SCF and IL - 6 cytokines for 48 h. The changes of CD49d (VLA -4) , CD11a ( LFA - 1 ) , CD62L ( L - selectin) and CD184 (CXCR4) were analyzed by flow cytometry. The adherent and migration activities of CD34^+ cells were evaluated in human fibronectin (FN) coated microplates (96 wells) and transwell system. RESULTS: The numbers of CD34^+ cell expanded to 3 folds and the percentages of CD34^+ cells that were positive expressions for CD49d, CDlla, CD62L or CD184 increased 1 to 2 folds after the cytokine stimulation. The spontaneous adhesion between CD34^+ , FN and SDF - 1 induced migration increased after SCF + IL -6 stimulated. CONCLUSION: SCF + IL -6 can improve the most of the homing related characteristics and activities in the short time expansion of CD34^+ hematopoietic stem/progenitor cells, which may be partly related to the increased intrinsic homing potential.
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