Lentiviral vectors efficiently transduce quiescent mature 3T3-L1 adipocytes
Lentiviral vectors efficiently transduce quiescent mature 3T3-L1 adipocytes
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DOI：
10.1016/j.ymthe.2003.11.021
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发表时间：
2004-02-01
影响因子：
12.4
通讯作者：
Hoeben, RC
中科院分区：
文献类型：
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作者：
Carlotti, F;Bazuine, M;Hoeben, RC
肥胖与许多严重的疾病有关，如心血管疾病、癌症和糖尿病。3T3-L1前体脂肪细胞分化模型是用于研究其潜在生理生物学过程的主要细胞系统之一。然而，对3T3-L1脂肪细胞的研究受到了这样一个事实的阻碍：众所周知，对成熟脂肪细胞进行基因改造是非常困难的。在这篇报道中，我们评估了慢病毒介导的基因转移到3T3-L1成熟脂肪细胞中的应用。我们证明了HIV-1来源的慢病毒载体可以有效地转导静止的、完全分化的3T3-L1脂肪细胞和3T3-L1前脂肪细胞。用LV-PGK-GFP慢病毒载体以每10(5)个细胞100 ng p24转导3T3-L1脂肪细胞后，95%以上的3T3-L1脂肪细胞表达报告基因。在培养中没有明显的毒性或细胞致病迹象。此外，对未分化前脂肪细胞的修饰不会影响其分化能力。此外，胰岛素诱导的葡萄糖摄取不受该程序的影响。相反，腺病毒介导的基因转移到3T3-L1脂肪细胞与显著的细胞致病性有关。根据这些数据，我们得出结论，慢病毒载体是成熟脂肪细胞基因修饰的首选基因转移系统。高效载体系统的可获得性可能会刺激脂肪组织作为肥胖症和其他疾病基因治疗的靶点。
Obesity is associated with many serious afflictions such as cardiovascular disease, cancer, and diabetes. One of the main cellular systems used to study the underlying physiological and biological processes is the 3T3-L1 preadipocyte differentiation model. However, studies on 3T3-L1 adipocytes are hampered by the fact that genetic modification of mature adipocytes is notoriously difficult. In this report, we evaluated the use of lentivirus-mediated gene transfer into 3T3-L1 mature adipocytes. We demonstrate that quiescent, fully differentiated 3T3-L1 adipocytes as well as 3T3-L1 preadipocytes can be efficiently transduced with HIV-1-derived lentiviral vectors. Upon transduction using LV-PGK-GFP lentiviral vector at 100 ng p24 per 10(5) cells, more than 95% of the 3T3-L1 adipocytes in the culture expressed the GFP reporter gene. There were no overt signs of toxicity or cytopathogenicity in the cultures. Furthermore, modification of undifferentiated preadipocytes did not affect their capacity to differentiate. In addition, insulin-induced glucose uptake was not affected by the procedure. In contrast, adenoviral-mediated gene transfer into 3T3-L1 adipocytes is associated with marked cytopathogenicity. From these data, we conclude that lentiviral vectors are the gene-transfer system of choice for genetic modification of mature adipocytes. The availability of an efficient vector system may stimulate the use of adipose tissue as a target for gene therapy in obesity and other disorders.