Pierelli L, Scambia G, Menichella G, Fattorossi A, Ciarli M, Bonanno G, Battaglia A, d'Onofrio G, Benedetti Panici P, Iacone A, Mancuso S, Leone G
Centro Ricerche per la Manipolazione dei Costituenti Ematici, Catholic University, Rome, Italy.
Br J Haematol. 1997 Jan;96(1):55-63. doi: 10.1046/j.1365-2141.1997.8632491.x.
A combination of erythropoietin (EPO) plus stem cell factor (SCF) drove purified unfractionated granulocyte colony stimulating factor (G-CSF)/chemotherapy mobilized peripheral blood CD34+ cells to selective erythroid differentiation in liquid culture with an average 28-fold increase in the total cell number after 21 d. From day 6 of culture cytologic and cytofluorimetric characterization revealed that cultured cells belonged to the erythroid lineage with a gradual wave of maturation along the erythroid pathway to terminal cells. A similar pattern of erythroid differentiation was observed when the same peripheral blood CD34+ cells were culture with EPO plus SCF in serum-free medium. This cytokine combination produced selective erythroid differentiation with the complete exhaustion of the clonogenic potential on day 21. In parallel experiments the same circulating CD34+ cells underwent granulocytic/ monocytic differentiation in liquid culture in response to granulocyte-macrophage colony stimulating factor (GM-CSF), interleukin-3 (IL-3) and SCF, demonstrating that these CD34+ progenitors had intact pluripotent differentiating potential. Conversely, bone marrow CD34+ cells isolated from bone marrow allografts were unable to selectively differentiate along the erythroid pathway when they were exposed to EPO plus SCF combination. However, these cells maintained a greater number of colony forming cells on day 21 of culture compared to mobilized peripheral blood CD34+ cells. This model is a simple and reliable way to obtain selective erythroid differentiation of peripheral blood G-CSF/ chemotherapy mobilized CD34+ progenitor cells in liquid culture. The absence of cytokines such as GM-CSF and IL-3 in the culture medium permits studies on in vitro erythropoiesis without disturbance of prevalent myelopoiesis.
促红细胞生成素（EPO）加干细胞因子（SCF）的组合，可促使经纯化的未分级粒细胞集落刺激因子（G-CSF）/化疗动员的外周血CD34+细胞在液体培养中选择性地向红系分化，21天后细胞总数平均增加28倍。从培养的第6天起，细胞学和细胞荧光分析表明，培养的细胞属于红系谱系，沿着红系途径逐渐成熟为终末细胞。当相同的外周血CD34+细胞在无血清培养基中与EPO加SCF一起培养时，观察到了类似的红系分化模式。这种细胞因子组合可产生选择性红系分化，在第21天时克隆形成潜能完全耗尽。在平行实验中，相同的循环CD34+细胞在液体培养中对粒细胞-巨噬细胞集落刺激因子（GM-CSF）、白细胞介素-3（IL-3）和SCF发生反应，进行粒系/单核系分化，表明这些CD34+祖细胞具有完整的多能分化潜能。相反，从骨髓同种异体移植物中分离的骨髓CD34+细胞在暴露于EPO加SCF组合时，无法选择性地沿红系途径分化。然而，与动员的外周血CD34+细胞相比，这些细胞在培养的第21天保持了更多的集落形成细胞。该模型是在液体培养中获得外周血G-CSF/化疗动员的CD34+祖细胞选择性红系分化的一种简单可靠的方法。培养基中不存在GM-CSF和IL-3等细胞因子，使得可以在不干扰普遍的髓系造血的情况下研究体外红细胞生成。
