Suhayla Alnajjar1,2, Ingo Nolte1, Jan Torben Schille1,2, Sina Sender2, Nares Trakoolju3, Simon Villa Perez2, Dietmar Zechner4, Brigitte Vollmar4, Christian Junghanss2, Hugo Murua Escobar5,6. 1. Small Animal Clinic, University of Veterinary Medicine Hannover, Hannover, Germany. 2. Division of Medicine, Haematology, Oncology and Palliative Medicine, University of Rostock, Rostock, Germany. 3. Research Institute for Farm Animal Biology (FBN), Institute of Genome Biology, Dummerstorf, Germany. 4. Institute for Experimental Surgery, University of Medicine Rostock, Rostock, Germany. 5. Division of Medicine, Haematology, Oncology and Palliative Medicine, University of Rostock, Rostock, Germany; Hugo.Murua.Escobar@med.uni-rostock.de. 6. Comprehensive Cancer Center MV, Campus Rostock, Rostock, Germany.
Abstract
BACKGROUND/AIM: Cancer cell inoculation is routinely used to evaluate novel therapeutic approaches in vivo. However, without reporter genes enabling deep tissue imaging, study of early tumor progression and therapeutic responses is often limited. We describe the establishment and characterization of two canine cancer cell lines stably expressing red fluorescence proteins as tools for later in vivo imaging. MATERIALS AND METHODS: Two red fluorescence cell lines were generated by plasmid transfection. Fluorescence protein expression was confirmed by flow cytometry and microscopy. Deep tissue imaging was demonstrated in mice using a NightOWL LB 983. Gene expression changes after transfection were analyzed by RNAseq. RESULTS: Both cell lines were detectable in vivo by subcutaneous injection of 1×106 cells. RNAseq revealed up to 2005 transfection-induced differentially expressed genes but no significant changes in cellular key pathways. CONCLUSION: The fluorescent cell lines provide a solid basis for future in vivo studies on canine cancer.
BACKGROUND/AIM: Cancer cell inoculation is routinely used to evaluate novel therapeutic approaches in vivo. However, without reporter genes enabling deep tissue imaging, study of early tumor progression and therapeutic responses is often limited. We describe the establishment and characterization of two canine cancer cell lines stably expressing red fluorescence proteins as tools for later in vivo imaging. MATERIALS AND METHODS: Two red fluorescence cell lines were generated by plasmid transfection. Fluorescence protein expression was confirmed by flow cytometry and microscopy. Deep tissue imaging was demonstrated in mice using a NightOWL LB 983. Gene expression changes after transfection were analyzed by RNAseq. RESULTS: Both cell lines were detectable in vivo by subcutaneous injection of 1×106 cells. RNAseq revealed up to 2005 transfection-induced differentially expressed genes but no significant changes in cellular key pathways. CONCLUSION: The fluorescent cell lines provide a solid basis for future in vivo studies on canine cancer.
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