Schattner M, Lefebvre P, Mingolelli S S, Goolsby C L, Rademaker A, White J G, Foster D, Green D, Cohen I
Atherosclerosis Program, Rehabilitation Institute of Chicago, Northwestern University, Illinois, USA.
Stem Cells. 1996 Mar;14(2):207-14. doi: 10.1002/stem.140207.
The effect of thrombopoietin (TPO) on megakaryocytopoiesis (MKP) has been mainly studied using clonogenic assays in murine systems. In this study, we evaluated MKP in liquid culture using human bone marrow cells. While interleukin 3 (IL-3) and stem cell factor (SCF) are potent activators of TPO-stimulated MKP in the murine system, only IL-3 exhibited synergistic activity with TPO in cultures of human bone marrow. The IL-3 effect on TPO-stimulated megakaryocyte (MK) proliferation, expressed as the absolute number of MKs per seeded CD34+ cell, was more pronounced with purified CD34+ cell (8 +/- 1.6 SE versus 2.8 +/- 0.7 SE in the presence and absence of IL-3, respectively) than with mononuclear cells (MNC) (16 +/- 2.8 SE versus 11 +/- 2.0 SE). This effect of IL-3 on TPO-stimulated MK proliferation was due to a general proliferation of all cell types since the relative frequency of MKs (32.1 +/- 3 SE and 55.8 +/- 3 SE in MNC and CD34+ cells, respectively) was not affected by IL-3. The effect of TPO alone, TPO + IL-3, TPO + SCF, and TPO + IL-3 + SCF on MK proliferation was examined in MNC and CD34+ cultures. Greater numbers of MK per seeded CD34+ were observed in MNC compared to CD34+ cultures under all conditions except when TPO was added with both IL-3 and SCF. The enhancing effect of MNC was also observed on MK ploidy in the presence of TPO and IL-3. While proliferation and ploidy increase with TPO concentration in the murine system, they are inversely related in the human system. A significant 2.5-fold enhancement of TPO-induced MK proliferation was observed when purified CD34+ cells were cultured in inserts separated from human bone marrow stroma, indicating that soluble stimulatory factors are released from the stroma. These observations will be useful for ex vivo expansion of MKs to treat post-transplant or chemotherapy-associated thrombocytopenia.
血小板生成素（TPO）对巨核细胞生成（MKP）的影响主要是在小鼠系统中使用克隆形成试验进行研究的。在本研究中，我们使用人骨髓细胞在液体培养中评估了MKP。虽然白细胞介素3（IL-3）和干细胞因子（SCF）在小鼠系统中是TPO刺激的MKP的有效激活剂，但在人骨髓培养物中只有IL-3与TPO表现出协同活性。IL-3对TPO刺激的巨核细胞（MK）增殖的影响，以每接种的CD34+细胞中MK的绝对数量表示，在纯化的CD34+细胞中（分别在有和没有IL-3的情况下为8±1.6 SE对2.8±0.7 SE）比在单核细胞（MNC）中（16±2.8 SE对11±2.0 SE）更明显。IL-3对TPO刺激的MK增殖的这种影响是由于所有细胞类型的普遍增殖，因为MK的相对频率（MNC和CD34+细胞中分别为32.1±3 SE和55.8±3 SE）不受IL-3影响。在MNC和CD34+培养物中检查了单独的TPO、TPO + IL-3、TPO + SCF和TPO + IL-3 + SCF对MK增殖的影响。在所有条件下，与CD34+培养物相比，在MNC中观察到每接种的CD34+有更多的MK，但当TPO与IL-3和SCF一起添加时除外。在TPO和IL-3存在的情况下，还观察到MNC对MK倍性的增强作用。在小鼠系统中，增殖和倍性随TPO浓度增加，而在人系统中它们呈负相关。当纯化的CD34+细胞在与人骨髓基质分离的插入物中培养时，观察到TPO诱导的MK增殖显著增强2.5倍，表明可溶性刺激因子从基质中释放。这些观察结果将有助于体外扩增MK以治疗移植后或化疗相关的血小板减少症。
