摘要
目的 研究造血因子白介素 11(IL 11)基因修饰的基质细胞对造血细胞体外扩增的影响。方法 采用逆转录病毒载体将IL 11基因转入基质细胞HFCL ,用Northernblot检测基质细胞HFCLIL 11基因的表达 ,用流式细胞仪检测IL 11基因修饰的HFCL支持的脐血CD3 4 + 造血细胞体外扩增中表型为CD3 4 + CD3 8- 早期祖细胞和表型为CD3 4 + CD41+ 巨核系定向祖细胞的比例。结果 基质细胞HFCL能够表达逆转录病毒介导的IL 11基因 ,并且在这种IL 11基因修饰的基质细胞支持下 ,脐血CD3 4 + 造血细胞经过7d扩增 ,扩增细胞中表型为CD3 4 + CD3 8- 的早期祖细胞和表型为CD3 4 + CD41+ 巨核系祖细胞的比例分别为 (1.62± 0 .2 3 ) %、(9.9±1.1) % ,高于未转基因HFCL的 (0 .8± 0 .2 3 ) %、(6.5± 1.8) % ,而在相同条件下细胞因子支持的扩增细胞中则分别为 (0 .19±0 .14 ) %、(6.0± 1.1) %。结论 基质细胞能够表达经逆转录病毒载体介导的IL 11基因 ,而且经IL 11基因修饰的基质细胞能显著促进CD3 4 + CD3 8- 的早期祖细胞和CD3 4 + CD41+ 巨核系祖细胞扩增。
Objective To investigate the effect of IL 11 gene modified stromal cells on ex vivo expansion of hematopoietic cells. Methods After IL 11 gene was transferred into stromal cell line HFCL with retroviral vector pLXSN, the expression of IL 11 mRNA was detected with Northern blot in HFCL IL 11 cells. The percentages of CD34 + CD38 - early progenitor and CD34 + CD41 + megakaryocyte committed progenitor were determined with flow cytometry after inoculation on stromal cell HFCL IL 11. Results The HFCL cells expressed IL 11 mediated by retroviral vector. In the expansion of CD34 + cells from cord blood inoculated on HFCL IL 11 cells for 7 d, the percentage of CD34 + CD38 - early progenitors was (1.62±0.23)% and that of CD34 + CD41 + megakaryocyte committed progenitors (9.9±1.1)%, both of which were higher than the expansion inoculated on untransferred HFCL cells [(0.80±0.23)%, (6.5±1.8)%, respectively] and higher than inoculated with cytokinases in the same conditions [(0.19±0.14)%, (6.0±1.1)%, respectively]. Conclusion Stromal cells can express the retroviral mediated IL 11. These HFCL IL 11 cells can enhance the expansion of CD34 + CD38 - early progenitor and CD34 + CD41 + megakaryocyte committed progenitors.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2001年第1期55-58,共4页
Journal of Third Military Medical University
基金
国家"973"重点基础研究项目!( 973) (G19990 5390 3)
国家杰出青年科学基金资助项目! ( 3982 5111)
