Labelling of human adipose-derived stem cells for non-invasive in vivo cell tracking
Labelling of human adipose-derived stem cells for non-invasive in vivo cell tracking
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DOI：
10.1007/s10561-006-9027-7
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发表时间：
2007-09-01
影响因子：
1.5
通讯作者：
Redl, Heinz
中科院分区：
文献类型：
--
作者：
Wolbank, Susanne;Peterbauer, Anja;Redl, Heinz
人脂肪源性干细胞（ASC）可以在未分化状态下扩增或沿成骨、成软骨、成脂、成肌、内皮和神经谱系沿着分化。为了测试它们的体内和原位再生潜力，需要在合适的缺损模型中应用后追踪它们的命运。非侵入性成像系统允许对活体动物中的标记细胞进行真实的时间跟踪。我们已经评估了一种生物发光细胞跟踪方法，以可视化在活体动物中用荧光素酶标记的ASC。已经测试了两种用报道基因（荧光素酶，绿色荧光蛋白）有效标记人干细胞的方法，即用LipofectamineTM 2000进行脂质转染和用Nucleofector装置进行电穿孔。使用脂质体转染和核转染方案，我们已经达到高达60%的转染效率。报告基因表达在体外3周内可检测到，并且不干扰细胞的表型和干细胞特性。通过高灵敏度的CCD照相机，我们能够实现细胞命运的真实的时间成像，在裸鼠中施用（静脉内、肌内、腹膜内、皮下）后至少20天。此外，我们能够通过选择不同的应用模式，如封闭在纤维蛋白基质中来影响细胞的流动性。具有瞬时转染的光学成像系统是一种优雅的细胞跟踪概念，用于跟踪小动物中人类干细胞的存活和命运。
Human adipose-derived stem cells (ASC) can be expanded in an undifferentiated state or differentiated along the osteogenic, chondrogenic, adipogenic, myogenic, endothelial and neurogenic lineage. To test their in vivo and in situ regenerative potential, their fate needs to be traced after application in suitable defect models. Non-invasive imaging systems allow for real time tracking of labelled cells in the living animal. We have evaluated a bioluminescence cell tracking approach to visualise ASC labelled with luciferase in the living animal. Two procedures have been tested to efficiently label human stem cells with a reporter gene ( luciferase, green fluorescent protein), namely lipofection with Lipofectamine (TM) 2000 and electroporation with a Nucleofector (R) device. With both lipofection and nucleofection protocols, we have reached transfection efficiencies up to 60%. Reporter gene expression was detectable for 3 weeks in vitro and did not interfere with the phenotype and the stem cell properties of the cells. By means of a highly sensitive CCD camera, we were able to achieve real time imaging of cell fate for at least 20 days after application ( intravenous, intramuscular, intraperitoneal, subcutaneous) in nude mice. Moreover, we were able to influence cell mobility by choosing different modes of application such as enclosure in fibrin matrix. The optical imaging system with transient transfection is an elegant cell-tracking concept to follow survival and fate of human stem cells in small animals.