title: Labelling of human adipose-derived stem cells for non-invasive in vivo cell tracking.
abstract: 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.
authors.affiliation: Red Cross Blood Transfusion Service of Upper Austria, Blumauerstr. 3-5, Linz, A-4020, Austria. susanne.wolbank@blutz.o.redcross.or.at
authors.forename: Susanne
authors.initials: S
authors.lastname: Wolbank
authors.forename: Anja
authors.initials: A
authors.lastname: Peterbauer
authors.forename: Esther
authors.initials: E
authors.lastname: Wassermann
authors.forename: Simone
authors.initials: S
authors.lastname: Hennerbichler
authors.forename: Regina
authors.initials: R
authors.lastname: Voglauer
authors.forename: Martijn
authors.initials: M
authors.lastname: van Griensven
authors.forename: Hans-Christoph
authors.initials: HC
authors.lastname: Duba
authors.forename: Christian
authors.initials: C
authors.lastname: Gabriel
authors.forename: Heinz
authors.initials: H
authors.lastname: Redl
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D002454:Cell Differentiation; D049109:Cell Proliferation; D003584:Cytological Techniques / Q000379:methods*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D010012:Osteogenesis; D013234:Stem Cells / Q000166:cytology*; D014162:Transfection; D051598:Whole Body Imaging
chemical_list: D049452:Green Fluorescent Proteins; D008156:Luciferases