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目的探讨不同细胞因子组合对脐带血中ACl33+细胞表面趋化因子受体(CXCR4)、粘附分子VLA4(CD49d)、LFA1(CD11a)和Lselectin(CD62L)表达的影响。方法采用直接免疫荧光标记法,使用流式细胞仪测定新鲜分离的、不同培养条件下体外培养7d、14d的脐带血中ACl33+细胞表面CXCR4和CD49d、CD11a和CD62L的表达情况。结果新鲜脐带血ACl33+细胞中CXCR4的表达率为(50.2±12.5)%;CD49d的表达率为(53.2±15.6)%,CD11a的表达率为(98.1±2.4)%,CD62L的表达率为(52.4±16.6)%。在无血清、无细胞因子存在的条件下,体外培养第7d及14d,CXCR4、VLA4、CD11a和CD62L的表达显著下降(P<0.05)。在有细胞因子存在的条件下,短期液体扩增培养后,脐带血ACl33+细胞中CXCR4、VLA4、CD11a和CD62L的表达率较新鲜ACl33+细胞均有不同程度的增高(P<0.05);IL3+FL+SCF组较IL11+FL3+SCF组表达率高,其中以培养第14dCXCR4、CD62L的表达增高最明显。结论在体外短期扩增培养过程中,早期效应细胞因子能显著上调脐带血ACl33+细胞CXCR4、CD49d、CD11a和CD62L等的表达;用SCF、FL和IL3组合扩增脐带血,不仅能获得足够数量的造血干/祖细胞,还能增加造血干/祖细胞的迁移和归巢能力。
Objective To investigate the effects of different combinations of cytokines on the expression of CXCR4, CD49d, CD11a and CD62L in cord blood of cord blood. Methods The expression of CXCR4, CD49d, CD11a and CD62L on ACl33 + cells in umbilical cord blood cultured freshly isolated and cultured in vitro for 7 days and 14 days were measured by direct immunofluorescence labeling. Results The positive rate of CXCR4 in fresh cord blood was (50.2 ± 12.5)%, that of CD49d was (53.2 ± 15.6)%, that of CD11a was (98.1 ± 2.4)% and that of CD62L ± 16.6)%. The expression of CXCR4, VLA4, CD11a and CD62L were significantly decreased (P <0.05) on the 7th day and the 14th day in vitro in the absence of serum and cytokines. In the presence of cytokines, the expression of CXCR4, VLA4, CD11a and CD62L in cord blood ACl33 + cells was increased to a greater extent than that of fresh ACl33 + cells (P <0.05) after short-term liquid amplification. IL3 + FL + SCF group was higher than that of IL11 + FL3 + SCF group, in which the expression of CXCR4 and CD62L increased most significantly on the 14th day. Conclusion In the short-term expansion of in vitro culture, early effector cytokines can significantly upregulate the expression of CXCR4, CD49d, CD11a and CD62L in cord blood ACl33 + cells. The combination of SCF, FL and IL3 can not only obtain a sufficient number of Hematopoietic stem / progenitor cells, but also increase the ability of hematopoietic stem / progenitor cells to migrate and homing.