title: Inhibition of oxygen-glucose deprivation-induced apoptosis of human adipose-derived stem cells by genetic modification with antiapoptotic protein bcl-2.
abstract: BACKGROUND
  Adipose-derived stem cells (ADSCs) have become a promising tool for a wide range of cell-based therapies. However, transplanted ADSCs do not survive well under ischemic conditions. In this study we aimed to inhibit oxygen-glucose deprivation (OGD)-induced apoptosis of human ADSCs by genetic modification with antiapoptotic protein Bcl-2.
  
  
  METHODS
  After isolation and culture, the phenotypes of human ADSCs at passage 3 were analyzed by flow cytometry. Then, genetic modification of ADSCs with Bcl-2 was carried out. Bcl-2 gene transfection was verified by Western blot analysis and multipotent differentiation properties were evaluated in Bcl-2-modified ADSCs (Bcl-2-ADSCs). Apoptosis was evaluated by a TUNEL assay under ischemic conditions induced by OGD. Apoptotic nuclei were also assessed and quantified by Hoechst staining.
  
  
  RESULTS
  The cultured ADSCs expressed stem cell-associated markers CD29, CD34, CD44, and CD90, but not fibroblast marker HLA-DR or hematopoietic stem cell marker CD133. The Bcl-2 gene was transferred into ADSCs efficiently, and Bcl-2-ADSCs differentiated into adipocytes, chondrocytes, and osteoblasts. In addition, Bcl-2 overexpression reduced the percentage of apoptotic Bcl-2-ADSCs by 38 % under OGD.
  
  
  CONCLUSIONS
  Our results indicate that Bcl-2 overexpression through gene transfection inhibits apoptosis of ADSCs under ischemic conditions.
  
  
  METHODS
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authors.affiliation: Department of Burns and Plastic Surgery, The Drum Tower Clinical Medical College, Nanjing Medical University, Nanjing, 210008, China.
authors.forename: Ziwei
authors.initials: Z
authors.lastname: Cui
authors.forename: Liangyun
authors.initials: L
authors.lastname: Shen
authors.forename: Yue
authors.initials: Y
authors.lastname: Lin
authors.forename: Shuqin
authors.initials: S
authors.lastname: Wang
authors.forename: Dongfeng
authors.initials: D
authors.lastname: Zheng
authors.forename: Qian
authors.initials: Q
authors.lastname: Tan
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000328:Adult; D017209:Apoptosis / Q000502:physiology*; D015415:Biomarkers / Q000378:metabolism; D015687:Cell Hypoxia / Q000502:physiology; D064987:Cell- and Tissue-Based Therapy / Q000379:methods*; D002478:Cells, Cultured; D005260:Female; D005434:Flow Cytometry; D005947:Glucose / Q000172:deficiency; D006801:Humans; D039902:Multipotent Stem Cells; D010100:Oxygen / Q000502:physiology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis / Q000378:metabolism*; D013234:Stem Cells / Q000378:metabolism / Q000502:physiology*; D014162:Transfection
chemical_list: D015415:Biomarkers; D019253:Proto-Oncogene Proteins c-bcl-2; D005947:Glucose; D010100:Oxygen