组织工程与重建外科杂志

组织工程与重建外科杂志 ›› 2010, Vol. 6 ›› Issue (6): 306-310.doi: 10.3969/j.issn.1673-0364.2010.06.002

• 论著 • 上一篇    下一篇

腺病毒介导HGF基因转染对人脂肪干细胞生物学特性影响的研究

吴一杰，王黔，穆大力，栾杰   

  1. 北京协和医学院整形外科医院
  • 发布日期:2020-07-23

Study on Biological Characteristics of Adipose Derived Stem Cells Transfected with HGF Gene by Adenovirus Vector

WU Yijie,WANG Qian,MU Dali,LUAN Jie   

  1. Plastic Surgery Hospital,Chinese Academy of Medical Sciences,Beijing 100144,China
  • Published:2020-07-23

摘要： 目的观察腺病毒介导的HGF基因转染对人脂肪干细胞(Human adipose-derived stem cells,hADSCs)生物学特性的影响。方法利用腺病毒介导HGF(MOI=200)转染hADSCs,检测转染组细胞目的基因HGF表达情况,并与未转染组细胞的形态、细胞生长曲线、细胞表面标志物、细胞周期和成脂、成骨诱导分化能力进行比较。结果 hADSCs转染24 h后,即可检测出HGF,并于48 h后达到高峰,1周后HGF表达逐渐降低。hADSCs转染后仍呈成纤维细胞样生长,其生长增殖能力与未转染hADSCs无明显差别;转染后的hADSCs仍表达CD44、CD90、CD49d、CD105,不表达CD45、CD34、CD31、CD106和STRO-1。转染组与未转染组细胞周期、成脂和成骨诱导分化能力无明显差别。结论腺病毒表达载体介导HGF基因转染hADSCs并不影响其生物学特性,是将hADSCs用于基因治疗的良好途径。

关键词: 人脂肪干细胞, 腺病毒表达载体, 基因转染, 生物学特性

Abstract: Objective To investigate the effect of HGF transfection by adenovirus vector(Ad-HGF) on biological characteristics of human adipose derived stem cells(hADSCs).Methods hADSCs was transfected by Ad-HGF in MOI=200.The expression of HGF gene was detected by ELISA.The differences of biological characteristics such as cell morphology,growth curve,surface markers,cell cycle,differentiation potential between hADSCs transfected by HGF group and hADSCs group were observed.Results The expression of HGF was detected in supernatant 24 hours post transfection.The peak of it presented at 48 hours post transfection and sustained till 1 week after transfection.The hADSCs transfected by Ad-HGF grows as fibroblast-like style.There were no morphology and cell viability changes observed between transfected group and control group.Both two groups were shown to be positive for the expression of CD44,CD90,CD49d and CD105.On the other hand,expression of CD45,CD34,CD31,CD106 and STRO-1 were negative.There was no significant difference between two groups in cell cycle,adipogenic and osteogenic differentiation potential were observed.Conclusion The biological characteristics of hADSCs are not affected by adenovirus transfection.Adenovirus transfection is an ideal way to integrate HGF gene to hADSCs.

Key words: Human adipose derived stem cells, Adenovirus vector, Gene transfection, Biological characteristics

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