Forced differentiation in vitro leads to stress-induced activation of DNA damage response in hiPSC-derived chondrocyte-like cells

Human induced pluripotent stem cells (hiPSCs) constitute an important breakthrough in regenerative medicine, particularly in orthopedics, where more effective treatments are urgently needed. Despite the promise of hiPSCs only limited data on in vitro chondrogenic differentiation of hiPSCs are available. Therefore,the global gene expression profile of chondrocyte-like cells derived from hiPSCs (ChiPS) were compared to the hiPSCs (GPCCi001-A cell line) and to two cell lines of mature chondrocytes: HC-402-05a (Merck Millipore, Germany) (HC) and primary articular cartilage chondrocytes (ACC). GPCCi001-A cells were generated from primary human dermal fibroblasts in reprogramming process using Stemcca-tetO lentiviral vector coding transcription factors responsible for pluripotency (Oct 4, Sox2, c-Myc and Klf4). GPCCi001-A cells express markers characteristic for pluripotent SCs and possess capacity to form embryoid bodies with the derivatives of three primary germ layers. The experiment was performed in triplicate. We performed global gene expression analysis using the Affymetrix platform—to better understand the processes directing cell fate during chondrogenic differentiation in vitro (hiPSC,HC and ACC served as a control). This series includes GPCCi001-A and ChIPS samples re-used from GSE106239.

人类诱导多能干细胞（human induced pluripotent stem cells，hiPSCs）在再生医学领域，尤其是在亟需更有效治疗手段的骨科领域，是一项重要突破。尽管此类细胞拥有广阔应用前景，但目前针对其体外软骨分化的相关研究数据仍十分有限。因此，本研究将源自人类诱导多能干细胞的软骨细胞样细胞（chondrocyte-like cells derived from hiPSCs，ChiPS）的全基因表达谱，分别与人类诱导多能干细胞（GPCCi001-A细胞系）以及两种成熟软骨细胞系：德国默克密理博（Merck Millipore）旗下的HC-402-05a细胞系（简称HC）及原代关节软骨细胞（primary articular cartilage chondrocytes，ACC）进行了对比。GPCCi001-A细胞系源自原代人真皮成纤维细胞，通过搭载多能性相关转录因子（Oct4、Sox2、c-Myc及Klf4）的Stemcca-tetO慢病毒载体进行重编程构建而成。该细胞系可表达多能干细胞（pluripotent stem cells，SCs）特异性标志物，并具备形成包含三胚层衍生物的拟胚体的能力。本实验设置三次生物学重复。我们采用Affymetrix平台开展全基因表达分析，以期更深入阐明体外软骨分化过程中调控细胞命运的分子机制（以hiPSC、HC及ACC作为对照样本）。本数据集包含源自GSE106239的GPCCi001-A及ChiPS样本。