Cui Ziwei, Shen Liangyun, Lin Yue, Wang Shuqin, Zheng Dongfeng, Tan Qian
Department of Burns and Plastic Surgery, The Drum Tower Clinical Medical College, Nanjing Medical University, Nanjing, 210008, China.
Aesthetic Plast Surg. 2014 Aug;38(4):779-87. doi: 10.1007/s00266-014-0354-y. Epub 2014 Jun 7.
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.
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.
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.
Our results indicate that Bcl-2 overexpression through gene transfection inhibits apoptosis of ADSCs under ischemic conditions.
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脂肪来源干细胞（ADSCs）已成为广泛的基于细胞的治疗的一种有前景的工具。然而，移植的ADSCs在缺血条件下存活不佳。在本研究中，我们旨在通过用抗凋亡蛋白Bcl-2进行基因改造来抑制氧糖剥夺（OGD）诱导的人ADSCs凋亡。
分离培养后，通过流式细胞术分析第3代人ADSCs的表型。然后，用Bcl-2对ADSCs进行基因改造。通过蛋白质免疫印迹分析验证Bcl-2基因转染，并在Bcl-2修饰的ADSCs（Bcl-2-ADSCs）中评估多能分化特性。在OGD诱导的缺血条件下，通过TUNEL试验评估凋亡。还通过Hoechst染色评估和定量凋亡细胞核。
培养的ADSCs表达干细胞相关标志物CD29、CD34、CD44和CD90，但不表达成纤维细胞标志物HLA-DR或造血干细胞标志物CD133。Bcl-2基因有效地转入ADSCs，且Bcl-2-ADSCs分化为脂肪细胞、软骨细胞和成骨细胞。此外，Bcl-2过表达使OGD条件下凋亡的Bcl-2-ADSCs百分比降低了38%。
我们的结果表明，通过基因转染使Bcl-2过表达可抑制缺血条件下ADSCs的凋亡。
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