Liehr Thomas, Kankel Stefanie, Buhl Eva Miriam, Schröder-Lange Sarah K, Weiskirchen Ralf
Institute of Human Genetics, Jena University Hospital, Friedrich Schiller University, D-07747 Jena, Germany.
Electron Microscopy Facility, Institute of Pathology, RWTH University Hospital Aachen, D-52074 Aachen, Germany.
Cells. 2024 Dec 29;14(1):21. doi: 10.3390/cells14010021.
The Rat-1 cell line was established as a subclone of the parental rat fibroblastoid line F2408, derived from Fisher 344 rat embryos. Rat-1 cells are widely used in various research fields, especially in cancer biology, to study the effects of oncogenes on cell proliferation. They are also crucial for investigating signal transduction pathways and play a key role in drug testing and pharmacological studies due to their rapid proliferation. Therefore, Rat-1 cells are an indispensable research tool. While some cytogenetic information on their basic chromosomal features is available, detailed genomic analyses, such as karyotype analysis, short tandem repeat (STR) profiling, and whole-genome sequencing, have not been thoroughly conducted. As a result, the genetic stability and potential variations in Rat-1 cells over extended culture periods are poorly understood. This lack of comprehensive genetic characterization can limit the interpretation of experimental results and requires caution when generalizing findings from studies using this cell line. In this study, we describe the genetic characterization of the Rat-1 cell line. We established a karyotype, performed multicolor fluorescence in situ hybridization (mFISH), identified chromosomal losses and gains, and defined an STR profile for Rat-1 with 31 species-specific markers. Interestingly, the chromosomal imbalances found in Rat-1 cells resemble those found in human epithelioid sarcoma or liposarcoma. Additionally, we analyzed the transcriptome of Rat-1 cells through mRNA sequencing (mRNA-Seq) using next-generation sequencing (NGS). Finally, typical features of these fibroblastic cells were determined using electron microscopy, Western blotting, and fluorescent phalloidin conjugates.
Rat-1细胞系是从源自Fisher 344大鼠胚胎的亲代大鼠成纤维样细胞系F2408中分离出的一个亚克隆。Rat-1细胞广泛应用于各种研究领域，尤其是癌症生物学领域，用于研究癌基因对细胞增殖的影响。由于其快速增殖，它们在研究信号转导途径方面也至关重要，并且在药物测试和药理学研究中发挥关键作用。因此，Rat-1细胞是一种不可或缺的研究工具。虽然已有一些关于其基本染色体特征的细胞遗传学信息，但尚未全面开展详细的基因组分析，如核型分析、短串联重复序列（STR）分型和全基因组测序。结果，对于Rat-1细胞在长时间培养过程中的遗传稳定性和潜在变异了解甚少。这种缺乏全面遗传特征描述的情况可能会限制实验结果的解释，并且在推广使用该细胞系的研究结果时需要谨慎。在本研究中，我们描述了Rat-1细胞系的遗传特征。我们建立了核型，进行了多色荧光原位杂交（mFISH），鉴定了染色体的缺失和增加，并使用31种物种特异性标记定义了Rat-1的STR图谱。有趣的是，在Rat-1细胞中发现的染色体失衡类似于在人类上皮样肉瘤或脂肪肉瘤中发现的情况。此外，我们通过使用下一代测序（NGS）的mRNA测序（mRNA-Seq）分析了Rat-1细胞的转录组。最后，使用电子显微镜、蛋白质免疫印迹和荧光鬼笔环肽缀合物确定了这些成纤维细胞的典型特征。
