骨髓间充质干细胞作为重组腺病毒白细胞介素10表达系统载体细胞的研究

探讨间充质于细胞(MSC)作为重组腺病毒白细胞介素10基因表达系统(Ad.rIL-10)的载体细胞的可行性。方法 应用密度梯度离心结合贴壁法原代培养,并于体外扩增出MSC,运用流式细胞仪对MSC进行鉴定。用ELISA法检测外源基因rIL-10在MSC中的表达情况。结果 原代培养扩增出的MSC高表达CD29,CD105和CD166,不表达CD45,其中第4代MSC生长状态良好,12h贴壁率达90％。腺病毒包装IL- 10可以高效感染MSC,感染48 h后IL-10表达增加(48 h Ad.rIL-10组136.2μg/L;对照组5.1μg/L),并可持续维持高表达2周以上。结论 含外源基因IL-10的MSC高效而持久的表达目的基因,尤其是第1代至第4代的MSC,是基因治疗良好的细胞载体。

Bone mesenchymal stem cell as a cellular vector for recombinant adenoviral expression of IL-10

Objective To explore the feasibility of using mesenchymal stem cell (MSC) as a cellular vector for recombinant adenoviral expression of IL-10.Methods Bone marrow-derived MSCs were obtained by primary culture using density gradient centrifugation and wall adherence, and were then subjected to in vitro proliferation. The expression of exogenous rIL- 10 in MSCs was examined with ELISA.Results The amplified MSCs culture showed high expressions of CD29, CD105 and CD166, but no expression of CD45. At the 4th passage, the MSCs grew well and 90％ appeared wall-adhering for 12 hours.The edenovirus containing recombinant IL- 10 (Ad.rIL- 10) was found to transfeet the MSCs with high efficiency. The expression of IL-10 started to increase at 48 hours after transfection ( 136.2 μg/L in the Ad.rIL10 group vs 5.1 μg/L in the controls), and remained at a high level for over 2 weeks. Conclusion MSCs tranfected with rIL- 10 recombinant adenovirus may show high and long-lasting expression of target gene,especially at passages 1 through 4, and therefore can be promising as a cellular vector for gene therapy.