Mao Ping, Xu Li, Mo Wen-Jian, Yin Yi, Xu Yan-Li, Lin Xiu-Mei
Department of Hematology, The Affiliated First Municipal People's Hospital of Guangzhou Medical College, Guangzhou 510182, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2006 Apr;14(2):318-21.
This study was purposed to explore the effect of different cytokine combinations on the expansion of the mononuclear cells drived from umbilical cord blood (CB) ex vivo and expression of CXCR4 and CD49d on CD34+ cells after expansion. Human fresh CB mononuclear cells were cultured in serum-free and stroma-free medium containing different combinations of cytokine for 7 days. At day o and 7, the total cells were counted, CD34+ cells and CD34+CXCR4+, CD34+CD49d+ cells were assayed by flow cytometry, and CFU were determined. According to the different combinations of cytokine, experiments were divided into four groups: control, SF group (SCF + FL), SFT group (SCF + FL + TPO) and SFT6 group (SCF + FL + TPO + IL-6). The results showed that the SF (SF group) combination supported only low expansion of total cells, CD34+ cells and CFU. The addition of TPO in SF group restored UCB stem/progenitors expansion to a higher level than that in SF group, while there was no difference between groups SFT and SFT6 (P > 0.05). The cytokine combinations in groups SF, SFT and SFT6 all could upregulate the expression levels of CD49d and CXCR4 on expanded cord blood CD34+ cells, but there were no significant differences between groups SF, SFT and SFT6 (P > 0.05). It is concluded that SCF + FL has no strong synergistic effects on primitive hematopoietic cells. TPO plays an important role in enhancing expansion of umbilical cord blood hematopoietic cells, while IL-6 only shows a neutral effect on it. SCF + FL + TPO combination not only promotes progenitor cells expansion but also upregulates the expression of CD49d and CXCR4 on CD34+ cells from cord blood.
本研究旨在探讨不同细胞因子组合对体外扩增脐血来源的单个核细胞的影响以及扩增后CD34+细胞上CXCR4和CD49d的表达。将人新鲜脐血单个核细胞在含有不同细胞因子组合的无血清、无基质培养基中培养7天。在第0天和第7天，计数总细胞数，通过流式细胞术检测CD34+细胞以及CD34+CXCR4+、CD34+CD49d+细胞，并测定集落形成单位（CFU）。根据细胞因子的不同组合，实验分为四组：对照组、SF组（SCF+FL）、SFT组（SCF+FL+TPO）和SFT6组（SCF+FL+TPO+IL-6）。结果显示，SF组合（SF组）仅支持总细胞、CD34+细胞和CFU的低水平扩增。SF组中添加TPO可将脐血干/祖细胞扩增恢复到高于SF组的水平，而SFT组和SFT6组之间无差异（P>0.05）。SF组、SFT组和SFT6组中的细胞因子组合均能上调扩增后脐血CD34+细胞上CD49d和CXCR4的表达水平，但SF组、SFT组和SFT6组之间无显著差异（P>0.05）。结论是，SCF+FL对原始造血细胞无强烈协同作用。TPO在增强脐血造血细胞扩增中起重要作用，而IL-6对其仅表现出中性作用。SCF+FL+TPO组合不仅促进祖细胞扩增，还上调脐血CD34+细胞上CD49d和CXCR4的表达。
