慢性粒细胞白血病患者骨髓间充质干细胞的分离培养及成脂分化

目的建立慢性粒细胞白血病（chronic myeloid leukemia,CML）患者骨髓间充质干细胞分离纯化培养方法,为慢性粒细胞白血病的后续研究提供实验基础。方法采用密度梯度离心联合贴壁筛选法分离纯化骨髓间充质干细胞（Bone marrowmesenchymal stem cells,BMSCs）,并用地塞米松、牛胰岛素、吲哚美辛、1-甲基-3-异丁基-黄嘌呤诱导BMSCs向脂肪细胞分化。油红O染色,倒置显微镜下观察结果。结果成功分离得到BMSCs,倒置显微镜下观察到BMSCs呈细而长的梭形生长,或多角形生长。诱导48h后出现脂滴,3周后油红O染色显示80%的BMSCs分化为脂肪细胞。结论可以从CML患者的骨髓中分离纯化出间充质干细胞,在体外诱导的条件下,间充质干细胞能向脂肪细胞分化。

Isolation,cultivation and adipogenisis differentiation of bone marrow mesenchymal stem cells from the patient with chronic myeloid leukemia

Objective To establish the method of isolation and cultivation of bone marrow mesenchymal stem cells（BMSCs） from the patient with chronic myeloid leukemia（CML）.Methods BMSCs were isolated from bone marrow and purified by density gradient centrifugation and cultured in vitro.Then BMSCs were induced into adipocytes with special medium containing insulin,1-methyl-3-isobutylxanthine,indomethine and dexamethasone.And finally,the adipocytes were identified with Oil Red O Staining.Results BMSCs grew as adherent cells.The morphology of BMSCs was observed with inverted microscope,cells showed slender fiber or cerioid figure.Forty-eight hours after induction,some of the cells displayed accumulation of lipid droplets.Twenty-one days after induction,about 80% of the cells appeared to be adipocytes,as detected by Oil Red O staining.Conclusions BMSCs were isolated and cultured successfully from the patient with CML.They could be induced into adipocytes in vitro by using medium containing insulin,1-methyl-3-isobutylxanthine,indomethine and dexamethasone.