不同细胞因子组合对c-kit(上标 +)Lin(上标 -)细胞的体外扩增研究

[Objective] To explore the regulation and the best time of c-kit(superscript +)Lin(superscript -) cell proliferation from mouse bone marrow by various combinations of cytokines without serum and matrix. [Methods] c-kit(superscript +)Lin(superscript -) cells were isolated from mouse bone marrow by using a high-gradient magnetic cell sorting system (MACS) and expanded for 14 days under different combinations of stem cell factor(SCF), FLt-3 ligand (FL), thrombopoietin (TPO), hepatic growth factor (HGF). At the different time points total cells and c-kit(superscript +)Lin(superscript -) cells were counted. FTTC-Annexin-V was applied to detect apoptosis rate on the tenth day in each group. [Results] Cytokines in each group could expand c-kit(superscript +)Lin(superscript -) cells 3-19 times as many as before with the combination of SCE, FL, TPO, IL-3 and HGF. While IL-3 or HGF were added to the combination of SCF, FL and TPO, c-kit(superscript +)Lin(superscript -) cells were increased significantly, and the apoptosis rate decreased. When the two cytokines were added together, they showed synergistic actions with SCF, FL and TPO to expand c-kit(superscript +)Lin(superscript -) cells. SCF+FL+TPO+IL-3+HGF expanded c-kit(superscript +)Lin(superscript -) cells most obviously on the tenth day and An-nexin-V positive cells were only 6.59%. [Conclusion] SCF+FL+TPO+HGF+IL-3 show a stronger potential for expanding c-kit(superscript +)Lin(superscript -) cells in vitro without serum and matrix. And the best time to expand the cells is ten days.