流式细胞仪在小鼠造血干细胞基因转移效率测定中的应用

目的探讨流式细胞仪在造血干细胞基因转移效率测定中的应用价值。方法以干细胞因子（ＳＣＦ）、白细胞介素（ＩＬ）３、ＩＬ６单独或联合作用（ＳＣＦ／ＩＬ３、ＳＣＦ／ＩＬ６、ＩＬ３／ＩＬ６、ＳＣＦ／ＩＬ３／ＩＬ６）后的Ｂａｌｂ／ｃ小鼠造血干细胞作为靶细胞，应用含有绿色荧光蛋白（ＧＦＰ）基因的逆转录病毒载体ＧＣＧＰＸＳＮ介导的基因转移技术实施基因转移，采用ＦＡＣＳ型流式细胞仪测定基因转移效率。结果病毒上清转染的阳性对照组基因转移率为０．０６％，以不同细胞因子作用后基因转移效率提高至００７％～０．２０％，细胞因子作用前后比较，除ＳＣＦ／ＩＬ６组外，差异均有显著或非常显著意义（Ｐ＜００５或Ｐ＜００１）。其中以ＳＣＦ／ＩＬ３组与ＳＣＦ／ＩＬ３／ＩＬ６组的提高最为明显，均达０２０％，与其他各细胞因子组比较，差异也有非常显著意义（Ｐ＜００１）。结论应用流式细胞仪可测定出ＳＣＦ、ＩＬ３、ＩＬ６单独或联合作用对小鼠造血干细胞基因转移效率的不同影响，具有快速、方便、准确的优点。

Application of Flow Cytometry in Determination of the Efficiency of Gene Transfer into Mouse Hematopoietic Stem Cells

Objecctive To explore the value of flow cytometry technique in determination of gene transfer efficiency. Methods Balb/c mouse hematopoietic stem cells were treated with SCF, IL3,IL6 separately or in combination. Then gene transfer mediated by retroviral vector containing GFP and Neo gene was made. The efficiency of gene transfer was determined by flow cytometry. Results Gene transfer efficiency of positive control group is 0.06%. The experiment groups using some cytokine may rise to 0070%. There is a significant difference ( P <005 or PZ<001) between the experiment groups except SCF/IL6 group and positive control group. SCF/IL3 group and SCF/IL3/IL6 group may enhance gene transfer efficiency to 0.20%, showing a very significant difference ( PBZ<001) comparison with other experiment groups. Conclusions The use of cytokine may enhance gene transfer efficiency of mouse hematopoietic stem cells. Flow cytometry has many advantages than other methods, such as fast, convenient performance and accurat