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The Effects of Adenoviral Transfection of the Keratinocyte Growth Factor Gene on Epidermal Stem Cells: an In Vitro Study

机译:腺病毒转染角质形成细胞生长因子基因对表皮干细胞的体外研究

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摘要

Epidermal stem cells (ESCs) are characterized as slow-cycling, multi-potent, and self-renewing cells that not only maintain somatic homeostasis but also participate in tissue regeneration and repair. To examine the feasibility of adenoviral vector-mediated keratinocyte growth factor (KGF) gene transfer into in vitro-expanded ESCs, ESCs were isolated from samples of human skin, cultured in vitro, and then transfected with recombinant adenovirus (Ad) carrying the human KGF gene (AdKGF) or green fluorescent protein gene (AdGFP). The effects of KGF gene transfer on cell proliferation, cell cycle arrest, cell surface antigen phenotype, and β-catenin expression were investigated. Compared to ESCs transfected with AdGFP, AdKGF-transfected ESCs grew well, maintained a high proliferative capacity in keratinocyte serum-free medium, and expressed high levels of β-catenin. AdKGF infection increased the number of ESCs in the G0/G1 phase and promoted ESCs entry into the G2/M phase, but had no effect on cell surface antigen phenotype (CD49f+/CD71). The results suggest that KGF gene transfer can stimulate ESCs to grow and undergo cell division, which can be applied to enhance cutaneous wound healing.
机译:表皮干细胞(ESC)具有慢循环,多能和自我更新的特征,不仅维持体细胞体内稳态,而且还参与组织再生和修复。为了检查腺病毒载体介导的角质形成细胞生长因子(KGF)基因转移到体外扩增的ESC中的可行性,从人皮肤样本中分离出ESC,进行体外培养,然后用携带人KGF的重组腺病毒(Ad)转染基因(AdKGF)或绿色荧光蛋白基因(AdGFP)。研究了KGF基因转移对细胞增殖,细胞周期停滞,细胞表面抗原表型和β-catenin表达的影响。与转染AdGFP的ESC相比,转染AdKGF的ESC生长良好,在无角化细胞无血清培养基中保持高增殖能力,并表达高水平的β-catenin。 AdKGF感染增加了G0 / G1期的ESC数量并促进ESC进入G2 / M期,但对细胞表面抗原表型没有影响(CD49f + / CD71 -< / sup>)。结果表明,KGF基因转移可以刺激ESC的生长并发生细胞分裂,可用于增强皮肤伤口的愈合。

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