LaIuppa J A, Papoutsakis E T, Miller W M
Northwestern University Department of Chemical Engineering, Evanston, Illinois 60208-3120, USA.
Stem Cells. 1997;15(3):198-206. doi: 10.1002/stem.150198.
The goal of our study was to identify cytokine combinations that would result in simultaneous ex vivo expansion of both the megakaryocyte (Mk) and granulocyte lineages, since these cell types have the potential to reduce the periods of thrombocytopenia and neutropenia following chemotherapy. We investigated the effects of cytokine combinations on expansion of the Mk (CD41a+ cells and colony forming unit [CFU]-Mk) and granulocyte (CD15+ cells and CFU-granulocyte/monocyte [GM]) lineages. Peripheral blood CD34+ cells were cultured in serum-free medium with interleukin 3 (IL-3), stem cell factor (SCF), and various combinations of thrombopoietin (TPO), IL-6, GM-CSF, and/or G-CSF. The Mk lineage was primarily influenced by TPO in our cultures, although Mk and CFU-Mk numbers were increased when TPO was combined with IL-6. The primary stimulator of the granulocyte lineage was G-CSF, although many synergistic and additive effects were observed with addition of other factors. Expansion of CFU-GM increased upon addition of more cytokines. The cytokine combination of IL-3, SCF, TPO, IL-6, GM-CSF and G-CSF produced the greatest number of granulocytes and CFU-GM. The minimum cytokines necessary for expansion of both the Mk and granulocyte lineages included TPO and G-CSF, since no other factors examined could increase Mk and granulocyte numbers to the same extent. The number of hematopoietic progenitors produced in our culture system should be sufficient for successful engraftment following myelosuppressive therapy if produced on a scale of about one liter.
我们研究的目的是确定能够在体外同时扩增巨核细胞（Mk）和粒细胞谱系的细胞因子组合，因为这些细胞类型有可能缩短化疗后血小板减少和中性粒细胞减少的持续时间。我们研究了细胞因子组合对Mk（CD41a+细胞和集落形成单位[CFU]-Mk）和粒细胞（CD15+细胞和CFU-粒细胞/单核细胞[GM]）谱系扩增的影响。外周血CD34+细胞在无血清培养基中与白细胞介素3（IL-3）、干细胞因子（SCF）以及血小板生成素（TPO）、IL-6、粒细胞-巨噬细胞集落刺激因子（GM-CSF）和/或粒细胞集落刺激因子（G-CSF）的各种组合一起培养。在我们的培养体系中，Mk谱系主要受TPO影响，不过当TPO与IL-6联合使用时，Mk和CFU-Mk的数量会增加。粒细胞谱系的主要刺激因子是G-CSF，不过添加其他因子时观察到了许多协同和相加效应。添加更多细胞因子后CFU-GM的扩增增加。IL-3、SCF、TPO、IL-6、GM-CSF和G-CSF的细胞因子组合产生的粒细胞和CFU-GM数量最多。扩增Mk和粒细胞谱系所需的最少细胞因子包括TPO和G-CSF，因为所检测的其他因子都无法将Mk和粒细胞数量增加到相同程度。如果以约一升的规模生产，我们培养体系中产生的造血祖细胞数量应足以在骨髓抑制治疗后成功植入。
