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目的:借助质粒将增强型绿色荧光蛋白(EGFP)基因转染猪骨髓干细胞(MSC),观察转染后EGFP在(MSC)内表达情况及MSC生长特性。方法:1月龄小型香猪为MSC供体,EGFP基因转染MSC,计算转染效率,流式细胞仪鉴定G418筛选后EGFP基因稳定表达率和转染后细胞增殖周期改变。结果:转染率为(36.9±3.4)%;G418筛选后EGFP表达率75.6%,EGFP转染MSC G0/G1期MSC比例较对照组明显增加(P<0.01),S+G2/M期MSC比例减少(P<0.01)。结论:猪MSC经基因转染和G418筛选,获得EGFP稳定表达;转染和G418筛选后MSC增殖能力降低,G0/G1期MSC比例增加,S+G2/M期MSC比例下降。
OBJECTIVE: To transfect enhanced green fluorescent protein (EGFP) gene into porcine bone marrow stem cells (MSC) with plasmid and observe the expression of EGFP in (MSC) and MSC growth characteristics after transfection. Methods: One-month-old miniature piglets were served as MSC donors. EGFP gene was transfected into MSCs to calculate the transfection efficiency. The stable expression of EGFP gene after G418 selection was determined by flow cytometry and the change of cell cycle after transfection. Results: The transfection rate was (36.9 ± 3.4)%. The EGFP expression rate was 75.6% after G418 screening. The proportion of MSC in G0 / G1 phase transfected with EGFP was significantly higher than that in control group (P <0.01) The proportion decreased (P <0.01). CONCLUSION: Transfection of porcine MSC and selection by G418 results in the stable expression of EGFP. After transfection and G418 screening, the proliferative ability of MSC is decreased, the proportion of MSC in G0 / G1 phases is increased and the proportion of MSCs in S phase G2 / M phase is decreased.