Wolbank Susanne, Peterbauer Anja, Wassermann Esther, Hennerbichler Simone, Voglauer Regina, van Griensven Martijn, Duba Hans-Christoph, Gabriel Christian, Redl Heinz
Red Cross Blood Transfusion Service of Upper Austria, Blumauerstr. 3-5, Linz, A-4020, Austria.
Cell Tissue Bank. 2007;8(3):163-77. doi: 10.1007/s10561-006-9027-7. Epub 2006 Oct 25.
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 2000 and electroporation with a Nucleofector 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.
人脂肪来源干细胞（ASC）可以在未分化状态下扩增，也可以沿成骨、成软骨、成脂、成肌、内皮和神经源性谱系分化。为了测试它们在体内和原位的再生潜力，在将其应用于合适的缺损模型后，需要追踪它们的命运。非侵入性成像系统能够对活体动物体内标记的细胞进行实时追踪。我们评估了一种生物发光细胞追踪方法，以在活体动物中可视化用荧光素酶标记的ASC。已经测试了两种用报告基因（荧光素酶、绿色荧光蛋白）有效标记人干细胞的方法，即使用Lipofectamine 2000进行脂质体转染和使用Nucleofector设备进行电穿孔。通过脂质体转染和核转染方案，我们都达到了高达60%的转染效率。报告基因表达在体外可检测3周，且不干扰细胞的表型和干细胞特性。通过高灵敏度的电荷耦合器件相机，在裸鼠中应用（静脉内、肌肉内、腹腔内、皮下）后，我们能够对细胞命运进行至少20天的实时成像。此外，通过选择不同的应用方式，如包封在纤维蛋白基质中，我们能够影响细胞迁移。具有瞬时转染功能的光学成像系统是一种精巧的细胞追踪概念，可用于追踪小动物体内人干细胞的存活和命运。
