目的:观察以质粒为载体,经脂质体介导的绿色荧光蛋白(greenfluorescent protein,GFP)转染的骨髓 间充质干细胞体外表达情况。 方法:以质粒病毒为载体,PT67细胞为包装细胞,介导绿色荧光蛋白转染新西兰 大白兔,在相差显微镜和荧光显微镜下观察,流式细胞仪检测GFP的表达效率。 结果:骨髓基质干细胞体外培 养后贴壁、集落生长,连续传代培养生长良好。 培养2wk后骨髓基质干细胞表面CD44抗原表达阳性。 介导绿 色荧光蛋白转染新西兰大白兔的骨髓基质干细胞,转染率可达30%左右。 结论:骨髓基质干细胞容易获取、来 源充足、可以大量扩增,可作为骨组织工程的种子细胞。 GFP基因的导入不会影响骨髓间充质干细胞的活性和 生物学特性。

Objective:Toobservetheexpressionof bonemarrowmesenchymal stemcells(BMSCs) invitro,whichis carriedby plasmidandtransfectedby greenfluorescent protein(GFP). Methods: Withplasmidvirus as thecarrier andPT67cells as thepackagingcells,mediatingGFPtotrasfect the NewZealand rabbits. Cells were observed under the phase contrast microscope and fluorescence microscope. GFPexpressionefficiencywas detectedbyflowcytometryinstrument. Results:BMSCs were colonial growthandevenamplificatedwell incontinuous passage culture. Additionally,CD44 onthe surface of BMSCs positively expressed after 2 weeks cultured in vitro. Detected by flowcytometry instrument,the transfection efficiency of GFP was above 30%. Conclusion:BMSCs can be easily obtainedandproliferated,therefore,theymaybeapplicatedas seed-cells inosseous tissueengineering. However,GFPgenewill not affect theactivityandbiological characteristics of BMSCs.

R392.4