移植物体外净化白血病细胞模型的建立

Establishment of Models for Purging Leukemic Cells from the Grafts In Vitro

本实验采用荧光素Calcein AM标记白血病细胞后分别接种入细胞株或骨髓细胞中以建立 3个白血病移植物细胞模型 ,采用免疫磁珠法对细胞模型进行选择 ,并用流式细胞术 (FCM)评价移植物模型中白血病细胞的净化效果。结果表明 :所建立的移植物细胞模型既可以用荧光显微镜分析 ,又可以用FCM进行精确评价。模型Ⅱ经免疫磁珠法体外净化后髓系白血病KG1a细胞的清除率为 (0 98± 0 0 9)log,模型Ⅲ中淋巴系白血病NALM 6细胞的清除率为 (1 82± 0 51 )log。结论 :建立的模型直观、稳定、重复性好 。

Using a fluorochrome Calcein AM, leukemia cells were labeled and seeded into cell lines or bone marrow cells to establish three cell models of grafts with leukemia. These cell models were engaged with CD34 immunomagnetic beads and the purging efficacy was evaluated using both fluorescence microscopy and flow cytometry. The results showed that the cell models established in this study could be evaluated successfully not only with fluorescence microscopy but also flow cytometry. After CD34 positive selection, KG1a cells were removed by (0 98±0 09)log in model Ⅱ and NALM 6 cells were rewosed by (1 82±0 51)log in model Ⅲ, respectively. It is concluded that the models established in this study are stable and direct with an excellent reproducibility and an accuracy which can be used to evaluate purging efficacy of leukemia cells in model graft using immunomagnetic selection and the experimental studies on tumorcidal effect in vitro.