Tpo、IL-11基因修饰的基质细胞对CD_(34)^+CD_(38)^-早期造血祖细胞扩增的影响

Effect of Tpo and/or IL-11 gene modified stromal cells on the expansion of CD_(34)^+CD_(38)^- hematopoietic primitive progenitor cells

目的 观察经Tpo、IL 11基因修饰的基质细胞对脐血CD3 4 + CD3 8-细胞体外扩增的影响。方法 用载有Tpo、IL 11基因的重组逆转录病毒感染成纤维样基质细胞HFCL ,通过Northernblot法检测基因修饰的HFCL细胞Tpo、IL 11基因的表达。以未经基因修饰的HFCL细胞作为对照 ,将脐血CD3 4 + 造血干 /祖细胞在这种基因修饰的HFCL细胞支持下 ,进行 7d体外扩增后 ,用锥虫蓝拒染法计数活细胞总数 ,并用流式细胞仪分析扩增细胞中CD3 4 + 细胞以及CD3 4 + CD3 8-细胞的比例。结果 在Tpo基因修饰的HFCL细胞、IL 11基因修饰的HFCL细胞和Tpo +IL 11基因共同修饰的HFCL细胞的支持下 ,扩增后CD3 4 + CD3 8-早期造血祖细胞的比例分别为 (1.8± 0 .2 4 ) %、(1.6 2± 0 .2 3) %、(2 .4 5±0 2 8) % ,细胞扩增倍数为 4 .2倍、3.6倍、6 .9倍 ,高于对照组的 (0 .80± 0 .2 3) %和 1.5倍 ,同时扩增细胞总数和CD3 4 + 细胞比例亦高于未经基因修饰的HFCL细胞所支持的扩增体系。结论 Tpo、IL 11基因修饰的基质细胞可有效促进脐血CD3 4 + 造血干 /祖细胞体外扩增 ,同时能有效维持扩增体系中的CD3 4 + CD3 8-细胞以及促进其扩增。

Objective To investigate the effects of Tpo and/or IL-11 gene modified stromal cells on the expansion of CD_ 34 + hematopoietic stem/progenitor cells in cord blood. Methods Retroviral vectors containing Tpo or IL-11 gene were constructed and used to transfect the stromal cell line HFCL. Tpo and/or IL-11 mRNA was assayed by Northern blot. Non-modified stromal cells were used, CD_ 34 + hematopoietic stem/progenitor cells from cord blood were expanded on gene-modified stromal cells for 7 days. The phenotype of CD_ 34 +CD_ 38 - primitive progenitors was detected by flow cytometry. Results HFCL expressed Tpo and/or IL-11 mRNA after transfected by the retroviral vectors. The percentages of CD_ 34 +CD_ 38 - primitive progenitors in the cultures of Tpo, IL-11 and Tpo+IL-11 modified HFCL were (1.8±0.24)%, (1.62±0.23)%, and (2.45±0.28)%, respectively, which were higher than that in the control [(0.8±0.23)%]. Conclusion The stromal cells modified by Tpo and/or IL-11 gene were able to enhance ex vivo expansion of CD_ 34 + and CD_ 34 +CD_ 38 - hematopoietic stem/progenitor cells from cord blood.