BACKGROUND:To improve the survival rate of transplanted tissue, most scholars focus on cel therapy, particularly cel-assisted fat grafting. OBJECTIVE:To analyze the effect of recombinant adenovirus-mediated hypoxia inducible factor 1alpha (HIF1α) mutant on survival rate of transplanted fat particles through transfection of adipose-derived mesenchymal stem cels. METHODS:Recombinant adenovirus-mediatedtriple-mutantHIF1αwas inserted into an adenovirus pAdEasy-1 system, folowed by viral packaging and titer determination. Human adipose-derived mesenchymal stem cels were cultured, passaged and identified, and subsequently transfected with three kinds of viruses and blank vector (experimental group with transfection of the triple mutantof HIF1α;positive control group; negative control group; blank control group). Transfection efficiency was determined using enhanced green fluorescent protein labeling. Additionaly, MTT assay was used to detect cel proliferation. RESULTS AND CONCLUSION:The recombinant adenovirus was successfuly constructed and packaged in line with transfection requirements. Moreover, adipose-derived mesenchymal stem cels were successfuly identified by adipogenic, osteogenic and chondrogenic induction and could be used asseed cels for subsequent experiments. RT-PCR results showed that HIF1α mRNA expression in the experimental group and positive control group was significantly higher than that in the other two groups (P< 0.05). Western blot analysis showed that the relative absorbance value in the experimental group was significantly higher than that in the other three groups (P< 0.05). A significant increase in the cel proliferation was found in the experimental group, significantly different from the other three groups (P<0.05). Therefore, our findings indicate that transfection of adenovirus-mediated triple-mutant HIF1α not only can sustain the expression of target protein in transfected adipose-derived mesenchymal stem cels under normoxic conditions, but also can promote the proliferation of transfected adipose-derived mesenchymal stem cels.%背景：为了提高移植组织的存活率，多数学者把研究的焦点放在了细胞治疗上，特别是经细胞辅助的脂肪移植领域。 目的：分析三点突变型低氧诱导因子1α转染脂肪间充质干细胞后促进脂肪移植成活率的作用。 方法：①利用目的片段重组三点突变型低氧诱导因子1α基因入腺病毒 pAdEasy-1系统，包装病毒并测定滴度；②人脂肪间充质干细胞的培养传代及鉴定；③以3种病毒液连同空白组共分4组进行后续实验(实验组含三点突变型低氧诱导因子1α基因；阳性对照组；阴性对照组；空白组)；将病毒液转染入脂肪间充质干细胞内，通过示踪因子增强型绿色荧光蛋白观察病毒转染效率，检测各组细胞中低氧诱导因子1α基因mRNA和蛋白表达情况；④通过MTT法检测各组脂肪间充质干细胞的增殖情况。 结果与结论：①腺病毒重组体构建成功并包装成功，符合转染要求；②通过成脂诱导、成骨诱导及成软骨诱导鉴定证明脂肪间充质干细胞培养成功，可以作为后续实验的种子细胞；③RT-PCR 结果显示，实验组和阳性对照组细胞内低氧诱导因子1α mRNA表达量均明显高于另外2组(P<0.05)；④Western blot检测显示，实验组蛋白表达相对吸光度值明显高于其他3组(P <0.05)；⑤实验组转染脂肪间充质干细胞后分裂增殖程度显著增加，组间比较差异有显著性意义(P<0.05)；⑥结果提示腺病毒介导的三点突变型低氧诱导因子1α基因转染脂肪间充质干细胞后不仅可以在常氧条件下持续表达目的蛋白，同时也能促进脂肪间充质干细胞的显著增殖，达到一种双赢的效果。
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