Targeted RNA-seq (TempO-Seq) of human hepatocellular carcinoma cell line HepG2, primary human hepatocytes (PHHs) and human induced pluripotent stem cell derived hepatocyte like cells (HLCs) in untreated condition

We aimed to compare the baseline expression of liver function and metabolism-related genes across the three liver models. Liver in vitro models are commonly used to study liver biology, disease mechanisms, and drug metabolism. Understanding their baseline gene expression profiles is crucial for interpreting experimental results and selecting the most appropriate model for specific research questions. Experimental workflow Cell Culture: Culture HepG2, PHHs, and hiPSCs-derived HLCs under standard, untreated conditions. Cell lysis: Collect RNA lysates using BioSpyder 2x enhanced lysis buffer from each cell type. Sequencing: Ship lysates to BioClavis, 201 Dumbarton Rd, Glasgow G81 4XJ, United Kingdom and sequence using TempoSeq targeted sequencing technology: https://www.bioclavis.co.uk/temposeq Data Analysis: Perform quality control of the sequencing results, normalize the data and compare normalized gene expression profiles, focusing on liver function and metabolic genes across the three models.

本研究旨在对比三种肝脏模型中肝功能及代谢相关基因的基线表达水平。肝脏体外模型常被用于研究肝脏生物学特性、疾病发生机制及药物代谢过程。明确其基因表达基线谱，对解读实验结果、为特定研究问题选取最适配的模型至关重要。
实验流程
细胞培养：将HepG2细胞、原代人肝细胞（PHHs）以及诱导多能干细胞（hiPSCs）分化得到的肝样细胞（HLCs）置于标准无处理条件下培养。
细胞裂解：从每种细胞中使用BioSpyder 2x增强型裂解缓冲液收集RNA裂解液。
测序：将裂解液寄送至英国格拉斯哥市邓巴顿路201号（邮编G81 4XJ）的BioClavis公司，使用TempoSeq靶向测序技术进行测序：https://www.bioclavis.co.uk/temposeq
数据分析：对测序结果开展质量控制、数据标准化处理，并对比三种模型中肝功能及代谢相关基因的标准化基因表达谱。