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目的了解 G-CSF 对 bcr/abl~+-CD34~+细胞增殖、分化的影响。方法采集慢性粒细胞白血病(CML)患者的 ber/abl~+-CD34~+细胞,分别以0、10、100、1000 ng/ml 的 G-CSF 与之共培养,并以正常骨髓 CD34~+细胞为对照,通过锥虫蓝拒染法、流式细胞术和光学显微镜观察研究其细胞增殖、周期分布、抗原分化和形态变化特点。结果所有实验组 ber/abl~+-CD34~+细胞均明显增长,其中 G-CSF10 ng/ml 组培养48、96 h,其细胞数量著高于同期无 G-CSF 组(P<0.05);而正常 CD34~+细胞数只在G-CSF 存在的情况下增长明显,其中 G-CSF 100 ng/ml 组培养48、96、144 h 细胞数均显著高于同期无G-CSF 组(P 值分别为<0.05,0.01,0.01);G-CSF 10、100、1000 ng/ml 组的 ber/abl~+-CD34~+细胞培养144 h 其 G_0/G_1期比例显著低于 G-CSF 空白组(P<0.05);而正常 CD34~+细胞 G-CSF 10、100、1000ng/ml 组培养48和96 h 其 G_0/G_1期比例均明显低于无 G-CSF 组(P<0.01);bcr/abl~+-CD34~+细胞及正常 CD34~+细胞 CD34抗原表达均随培养时间延长而下降,伴随 CD33和 CD13抗原先升后降,其变化与 C-CSF 浓度无关,但 ber/abl~+-CD34~+细胞的各抗原分化显著快于正常 CD34~+细胞。ber/abl~+-CD34~+细胞和正常 CD34~+细胞均随着增殖分化表现出终末细胞的形态特征。结论 G-CSF 能促进ber/abl~+-CD34~+细胞及正常 CD34~+细胞的增殖,但并非前者增殖的必要条件。ber/abl~+-CD34~+细胞比正常 CD34~+细胞分化更快,但两类细胞的分化速度均与 G-CSF 浓度无关。
Objective To investigate the effect of G-CSF on the proliferation and differentiation of bcr / abl ~ + -CD34 ~ + cells. Methods Bcl-la / CD34 + cells from patients with chronic myelogenous leukemia (CML) were collected and co-cultured with G-CSF at 0, 10, 100 and 1000 ng / Cells were used as control. The cell proliferation, cycle distribution, antigen differentiation and morphological changes were observed by trypan blue exclusion assay, flow cytometry and light microscopy. Results The number of cells in all experimental groups was significantly increased (P <0.05). The number of cells in G-CSF at 10 ng / ml for 48 and 96 h was higher than that in G-CSF at the same period (P <0.05). However, the number of normal CD34 + cells increased only in the presence of G-CSF. The number of cells cultured in G-CSF 100 ng / ml for 48, 96 and 144 h were significantly higher than those in the non-G-CSF group (P <0.05,0.01,0.01 respectively). The proportion of G 0 / G 1 phase in ber / abl ~ + -CD34 ~ + cells cultured in G-CSF 10,100,1000 ng / ml for 144 h was significantly lower than that in G- (P <0.05). The percentage of G_0 / G_1 phase of G-CSF at 10, 100 and 1000ng / ml in normal CD34 + cells was significantly lower than that in non-G-CSF groups at 48 and 96 h CD34 + cells and normal CD34 + CD34 antigen expression decreased with the prolongation of culture time, with the CD33 and CD13 antigen first and then decreased, the change has nothing to do with the concentration of C-CSF, but the ratio of ber / abl ~ The differentiation of + CD34 + cells was significantly faster than that of normal CD34 + cells. The morphological characteristics of terminal cells were observed with the proliferation and differentiation of both ber / abl ~ + -CD34 ~ + cells and normal CD34 ~ + cells. Conclusion G-CSF can promote the proliferation of ber / abl ~ + -CD34 ~ + cells and normal CD34 ~ + cells, but not the prerequisite for the proliferation of the former. The differentiation of ber / abl ~ + -CD34 ~ + cells was faster than that of normal CD34 ~ + cells, but the differentiation rate of both cells was not related to the concentration of G-CSF.