急性髓系白血病患者来源白血病干细胞的分选与鉴定

Isolation and identification of leukemia stem cells from acute myeloid leukemia patients

目的 分选急性髓系白血病患者白血病干细胞并进行鉴定,为白血病干细胞的进一步研究奠定基础.方法 采用Ficoll密度梯度离心法从患者骨髓中分离单个核细胞.采用流式细胞术从单个核细胞中分选CD34^＋CD123^＋白血病干细胞,通过集落形成能力和鹅卵石形成能力检测分选后白血病干细胞的自我更新与分化能力,同时检测CD34^＋CD123^＋细胞纯度,观察其细胞形态.结果 分选后的CD34^＋CD123^＋白血病干细胞比例占总分选细胞的10.7%,具有集落形成能力和鹅卵石形成能力,CD34^＋CD123^＋细胞纯度为62.1%.结论 成功分选与鉴定了白血病干细胞,可用于其后的进一步研究工作.

Objective To isolate and identify leukemia stem cells from acute myeloid leukemia patients for further research. Methods By Ficoll density gradient centrifugation, mononuclear cells were firstly separated from bone marrow of patients. According to specific surface markers, CD34^＋CD123^＋ of leukemic stem cells were sorted by flow cytometer. Their ability of self-renewal and differentiation were evaluated by colony formation and cobblestone forming ability. At the same time, the purity and cell morphology of CD34^＋CD123^＋ cells was analysed. Results Comparared with total mononuclear cells, the proportion of the CD34^＋CD123^＋ cells after sorting was 10.7 %, and these cells showed the ability of colony forming and cobblestone forming, and the purity proportion of CD34^＋CD123^＋ cells was 62.1%. Conclusion The leukemia stem cells were isolated successfully and could be used in further study.