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目的分离慢性粒细胞白血病(ChronicMyelogenousLeukemia,CML)患者骨髓中表达BCR/ABL肿瘤基因的细胞亚群,并研究其生物学特性。方法淋巴细胞分离液分离CML患者骨髓单个核细胞,免疫磁珠分选CD45-、GlyA-及CD34-细胞,极限稀释法获得单克隆细胞,流式细胞术鉴定其免疫表型,体外定向诱导分化检测其干细胞特性,并检测其Bcr/Abl融合基因的表达情况。结果CML患者骨髓源单克隆扩增细胞不表达造血细胞和内皮细胞的特征性表型,但Bcr/Abl融合基因阳性;诱导后的细胞能同时向造血细胞及血管内皮细胞分化。结论Bcr/Abl融合基因的产生至少发生在比造血干细胞更为早期的血液血管干细胞水平上,即CML至少起源于血液血管干细胞。
Objective To isolate the cell subset expressing BCR / ABL tumor gene in the bone marrow of patients with chronic myelogenous leukemia (CML) and to study its biological characteristics. Methods Lymphocyte separation liquid was used to separate bone marrow mononuclear cells from CML patients. CD45-, GlyA- and CD34- cells were sorted by immunomagnetic beads. Monoclonal cells were obtained by limiting dilution. The immunophenotypes were identified by flow cytometry. The stem cell characteristics were detected and the expression of Bcr / Abl fusion gene was detected. Results The bone marrow-derived monoclonal cells in CML patients did not express the characteristic phenotype of hematopoietic cells and endothelial cells, but the Bcr / Abl fusion gene was positive. The induced cells could differentiate into hematopoietic cells and vascular endothelial cells at the same time. Conclusion The Bcr / Abl fusion gene is produced at least in the early stage of hematopoietic stem cells compared with hematopoietic stem cells. That is, CML originated from hematopoietic stem cells.