RGB marking facilitates multicolor clonal cell tracking

• Published in: Nature medicine Vol. 17; no. 4; pp. 504 - 509
• Main Authors: Weber, Kristoffer, Thomaschewski, Michael, Warlich, Michael, Volz, Tassilo, Cornils, Kerstin, Niebuhr, Birte, Täger, Maike, Lütgehetmann, Marc, Pollok, Jörg-Matthias, Stocking, Carol, Dandri, Maura, Benten, Daniel, Fehse, Boris
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.04.2011; Nature Publishing Group
• Subjects: 631/1647/1407; 631/1647/1888/2249; 631/326/596/2561; Animals; Biomedical and Life Sciences; Biomedicine; Cancer Research; Cell division; Cell growth; Cell Tracking - methods; Cells; Cells (biology); Clone Cells - cytology; Clone Cells - metabolism; Cloning; Color; Dilution; Fluorescence; Fluorescent proteins; Gene expression; Gene therapy; Genetic aspects; Genetic Vectors; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Health aspects; Hematopoietic stem cells; Hepatocytes; Hepatocytes - cytology; Hepatocytes - metabolism; In vivo methods and tests; Infectious Diseases; Liver Regeneration; Luminescent Proteins - genetics; Luminescent Proteins - metabolism; Management; Marking; Metabolic Diseases; Mice; Mice, Inbred C57BL; Mice, Inbred NOD; Mice, SCID; Mice, Transgenic; Molecular Medicine; Neoplasm Transplantation; Neurosciences; Optical properties; Pathology; Physiological aspects; Progenitor cells; Proteins; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Red Fluorescent Protein; Regeneration; Scientific method; Spleen; Stems; technical-report; Tissue engineering; Transduction, Genetic; Transplantation; Tumor cells; Tumor Cells, Cultured - metabolism; Tumor Cells, Cultured - pathology; Tumors; United States; Germany
• ISSN: 1078-8956; 1546-170X
• DOI: 10.1038/nm.2338

Kristoffer Weber et al . describe a new lentiviral vector-mediated RGB (red, green and blue) multicolor cell marking technique for analyzing clonal cell fates in vitro and in vivo . Here they use RGB marking to assess clonality after regeneration of injured livers by transplanted primary hepatocytes, to mark hematopoietic stem/progenitor cells and to assess the clonality of tumor cells. The approach can potentially be adapted to various cell types and other vector systems. 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.