慢性粒细胞白血病骨髓间充质干细胞的分离及其支持造血的研究

目的探讨慢性粒细胞白血病（CML）患者骨髓间充质干细胞（MSCs）的分离以及体外支持造血的能力。方法采用细胞贴壁法获取16例CML患者的骨髓MSCs，在低血清培养液中培养和扩增。流式细胞仪检测免疫表型。通过油红O染色和Von Kossa染色检测MSCs向脂肪和骨的分化。逆转录聚合酶链法（RTPCR）检测bcr／abl融合基因的表达。对MSCs进行核型分析。RT-PCR检测细胞因子表达情况。将CML患者的骨髓MSCs作为滋养层，应用甲基纤维素半固体培养，检测其支持造血能力。结果CML来源的MSCs在倒置显微镜下为梭形，表达CD29、CD44、CD105，而CD14，CD31、CD34、CD45、HLA-DR均为阴性。在相应的诱导条件下可以向骨和脂肪分化。CML来源的MSCs不表达bcr／abl融合基因，具有正常细胞核型。CML来源的MSCs表达造血相关因子，在体外可以维持和扩增造血干／祖细胞。结论CML患者骨髓中存在具有多向分化能力的MSCs，其具有在体外支持造血的功能。

Isolation and characterization of chronic myelogenous leukemia bone marrow mesenchymal stem cells and their hematopoiesis support ability

Objective To study the characteristics of mesenchymal stem cells(MSCs)derived from chronic myelogenous leukemia(CML)patients' bone marrow and examine their hematopoiesis support in vitro.Methods MSCs from CML were obtained and cultured.Immunophenotype and in vitro differentiation capacities were investigated.Moreover,the Ph chromosome and bcr/abl gene of CML derived MSCs were detected.The expression of cytokines was detected by RT-PCR,and hema- topoiesis support of MSCs in vitro was detected by long-term bone marrow culture and the methylcel- lulose progenitor assay.Results CML derived MSCs showed a typical fibroblast like morphology. They were positive for CD29,CD44 and CD105,while negative for CD14,CD31,CD45,CD34 and HLA-DR.Under suitable conditions,CML derived MSCs could differentiate into osteoblasts and adi- pocytes.Further,CML derived MSCs showed a normal choromosome,and did not express bcr/abl gene.At last,they expressed hematopoiesis cytokines and possessed ability of hematopoiesis support in vitro.Conclusion There are MSCs with multidirectional differentiation potential in CML bone mar- row and they possess the ability of hematopoiesis support.