Bcr/Abl融合基因阳性、Flk1+CD34-慢性粒细胞白血病细胞体内生物学特性的研究

目的体内研究慢性粒细胞白血病(CML)患者骨髓中BCR/ABL+、Flk1+CD34-细胞的白血病干细胞特性。方法取CML患者骨髓,淋巴细胞分离液分离单个核细胞,常规接种培养,免疫磁珠分选CD45-、GlyA-及CD34-的贴壁细胞,流式细胞术鉴定其免疫表型,将其输入经亚致死量照射的NOD/SCID小鼠体内,检测人源细胞的植入及分化情况。结果植入的CML患者骨髓源BCR/ABL+、Flk1+CD34-细胞在受体小鼠体内分化为白血病细胞,受体小鼠骨髓细胞中存在Bcr/Abl融合基因阳性细胞。结论Bcr/Abl融合基因阳性Flk1+CD34-细胞具有白血病干细胞特性。

The Primary Study of biological characteristics and isolation of Bcr/Abl +、Flk1 +CD34- stem cells of Chronic Myelogenous Leukemia

Objective To study the leukemia stem cell characteristics of the Flk1+CD34-cells carrying Bcr/Abl fusion gene from the bone marrow of CML patients in vivo. Methods Bone marrow was extracted from the Ph+ patients with CML, mononuclear cells were separated by a Ficoll-Paque gradient centrifugation, throw off the suspended cells, using magnetic-activated cell separation (MACS) harvested the CD45-, GlyA-, and CD34-from the adherent cells, then identified the immunophenotype by fluorescence-activated cell sorter (FACS), then transplanted the cells to the NOD/SCID mice, as well as detected the implantation and differentiation of the human origin cells, meanwhile observation the bone marrow hyperplasia of the receptor mice. Results The engrafted Flk1+ CD34-cells carrying Bcr/Abl fusion gene from the bone marrow of CML patients can differentiate to the leukemia cells, it can be detected the Bcr/Abl fusion gene positive cells from the bone marrow of receptor mice. Conclusion The Flk1+ CD34-cells carrying Bcr/Abl fusion gene have the characteristic of the leukemia stem cell.