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血小板第4因子对脐血CD34+细胞黏附功能的影响

Effect of platelet factor 4 on the adherence of cord blood CD34+ cells

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【作者】 卢士红冯义杨仁池刘拥军翟琼莉张志华韩忠朝

【Author】 LU Shi-hong, FENG Yi, YANG Ren-chi, LIU Yong-jun, ZHAI Qiong-li, ZHANG Zhi-hua, HAN Zhong-chao.State key Laboratory of Experimental Hematology, Institute of Hematology, CAMS & PUMC, Tianjin 300020, China

【机构】 中国医学科学院、中国协和医科大学血液学研究所实验血液学国家重点实验室实验血液学国家重点实验室 300020天津300020天津

【摘要】 目的 研究血小板第 4因子 (PF4 )对新鲜脐血CD34+ 细胞及扩增后脐血CD34+ 细胞黏附功能的影响 ;PF4对脐血CD34+ 细胞上的黏附分子CD4 9d及基质细胞趋化因子 (SDF 1)受体CXCR4的作用。方法 采用免疫磁珠法 (MACS)分选CD34+ 细胞 ,结晶紫染色测定细胞总黏附性 ,免疫荧光标记流式细胞仪测定CD4 9d及CXCR4的表达。结果 ①PF4可使新鲜脐血CD34 + 细胞总黏附性提高 ,且与剂量相关。②SDF 110 0ng ml可使脐血CD34+ 细胞总黏附性提高。③脐血CD34+ 细胞扩增 10d后未加PF4刺激的自发以及经SDF 1诱导的黏附功能开始下降 ,在扩增脐血CD34+ 细胞不同时间段加入 10 0ng mlPF4 ,脐血CD34 + 细胞对基质层的黏附能力始终保持较高水平 ,以 0天时脐血CD34 + 细胞黏附性为10 0 % ,扩增 14d时脐血CD34+ 细胞黏附性PF4组为 (2 6 2 .0 4± 6 4 .81) % ,同期对照组为 (6 4 .35±8 2 9) % ,经SDF 1诱导下扩增 14d的CD34+ 细胞的总黏附性PF4组为 (138.31± 32 .39) % ,同期对照组为 (6 7.6 6± 12 .4 4 ) %。④PF4 10 0ng ml作用于CD34 + 细胞时 ,CD4 9d 表达增长 13.0 2 % ,CXCR4表达增长17 33%。结论 PF4可使新鲜及扩增后的脐血CD34+ 细胞黏附功能增强 ,并促进CD4 9d及CXCR4的表达 ,提示PF4可能有助于脐血干细胞的归巢。

【Abstract】 Objective To investigate the effects of platelet factor 4(PF4) on the adherence, and the expressions of adherent molecules CD_ 49d and CXCR4 and the receptor of SDF-1 of fresh and expanded cord blood CD_ 34 + cells. Methods CD_ 34 + cells were isolated from cord blood using MACS immune magnetic beads. The adherent ability was assayed by using crystal violet staining and the expression of adherent molecule CD_ 49d and CXCR4 by FACS. Results ①PF4 could increase the adherent ability of the fresh cord blood CD_ 34 + cells, the effect being positively correlated with the dose of PF4. ②SDF-1 at concentration of 100?ng/ml increased the adherent ability of the fresh cord blood CD_ 34 + cells. ③The spontaneous and the SDF-1 induced adherent ability of the cord blood CD_ 34 + cells began to decrease after being cultured for 10 days without PF4, while in the presence of PF4 at 100 ng/ml,the ability of the cord blood CD_ 34 + cell adhering to the stroma layer still remained at higher level. At day 14,the adherent ability was (262.04±64.81)% and (64.35±8.29)% in PF4 group and control group, respectively,if it was defined as 100% at day 0. SDF-1 at concentration of 100 ng/ml induced adherent ability was (138.31±32.39)% and (67.66±12.44)% in PF4 group and control group, respectively. ④The expression of CD_ 49d and CXCR4 increased 13.02% and 17.33%, respectively, when incubated with PF4. Conclusions PF4 could increase the adherent ability and promote the expression of CD_ 49d and CXCR4 of the cord blood CD_ 34 + cells, suggesting that PF4 promote the circulating stem cells homing to the marrow in the process of stem cells transplantation.

【基金】 国家攀登计划项目 (95 专 10 ) ;天津市科技重大攻关资助项目 (0 0 3 114 3 11)
  • 【文献出处】 中华血液学杂志 ,Chinese Journal of Hematology , 编辑部邮箱 ,2003年09期
  • 【分类号】R33
  • 【被引频次】4
  • 【下载频次】79
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