Small Animal Clinic, University of Veterinary Medicine Hannover, Hannover, Germany.
Division of Medicine, Haematology, Oncology and Palliative Medicine, University of Rostock, Rostock, Germany.
In Vivo. 2022 Jan-Feb;36(1):170-179. doi: 10.21873/invivo.12688.
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.
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.
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.
The fluorescent cell lines provide a solid basis for future in vivo studies on canine cancer.
背景/目的：癌细胞接种通常用于在体内评估新的治疗方法。然而，如果没有能够进行深层组织成像的报告基因，早期肿瘤进展和治疗反应的研究往往受到限制。我们描述了两种稳定表达红色荧光蛋白的犬癌细胞系的建立和特征，作为以后体内成像的工具。
通过质粒转染生成了两种红色荧光细胞系。通过流式细胞术和显微镜确认荧光蛋白的表达。使用 NightOWL LB 983 在小鼠中进行深层组织成像。转染后通过 RNAseq 分析基因表达变化。
通过皮下注射 1×106 个细胞，两种细胞系均可在体内检测到。RNAseq 显示多达 2005 个转染诱导的差异表达基因，但细胞关键途径没有明显变化。
荧光细胞系为未来的犬癌体内研究提供了坚实的基础。
