title: RGB marking facilitates multicolor clonal cell tracking.
abstract: We simultaneously transduced cells with three lentiviral gene ontology (LeGO) vectors encoding red, green or blue fluorescent proteins. Individual cells were thereby marked by different combinations of inserted vectors, resulting in the generation of numerous mixed colors, a principle we named red-green-blue (RGB) marking. We show that lentiviral vector-mediated RGB marking remained stable after cell division, thus facilitating the analysis of clonal cell fates in vitro and in vivo. Particularly, we provide evidence that RGB marking allows assessment of clonality after regeneration of injured livers by transplanted primary hepatocytes. We also used RGB vectors to mark hematopoietic stem/progenitor cells that generated colored spleen colonies. Finally, based on limiting-dilution and serial transplantation assays with tumor cells, we found that clonal tumor cells retained their specific color-code over extensive periods of time. We conclude that RGB marking represents a useful tool for cell clonality studies in tissue regeneration and pathology.
authors: Research Department Cell and Gene Therapy, Clinic for Stem Cell Transplantation, University Cancer Center Hamburg, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors: Kristoffer
authors: K
authors: Weber
authors: Michael
authors: M
authors: Thomaschewski
authors: Michael
authors: M
authors: Warlich
authors: Tassilo
authors: T
authors: Volz
authors: Kerstin
authors: K
authors: Cornils
authors: Birte
authors: B
authors: Niebuhr
authors: Maike
authors: M
authors: Täger
authors: Marc
authors: M
authors: Lütgehetmann
authors: Jörg-Matthias
authors: JM
authors: Pollok
authors: Carol
authors: C
authors: Stocking
authors: Maura
authors: M
authors: Dandri
authors: Daniel
authors: D
authors: Benten
authors: Boris
authors: B
authors: Fehse
mesh_terms: D000818:Animals; D058948:Cell Tracking / Q000379:methods*; D002999:Clone Cells / Q000166:cytology* / Q000378:metabolism*; D003116:Color; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism; D008115:Liver Regeneration; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D008822:Mice, Transgenic; D009368:Neoplasm Transplantation; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014161:Transduction, Genetic; D014407:Tumor Cells, Cultured / Q000378:metabolism / Q000473:pathology; D000097573:Red Fluorescent Protein
chemical_list: D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D011994:Recombinant Proteins; D000097573:Red Fluorescent Protein