Replication Data for: Optimizing a three-dimensional spheroid clearing method for the imaging-based evaluation of cardiotoxicity

Background Toxicity evaluation based on two-dimensional cell culture shows differences from clinical results and has the disadvantage of not accurately reflecting cell-to-cell cross-signaling. Since almost all cells in the human body are arranged in a three-dimensional structure and constitute a tissue, the in vitro reproduction of a three-dimensional tissue composed of human cells can be used as an effective model for drug development and toxicity evaluation. The clearing technique improves image resolution and can implement three-dimensional bio-images throughout the organization, enabling more efficient toxicity evaluation of disease model analysis using spheroids. Methods We generate an 100-200um spheroids and optimized clearing condition. Next application and imaging of optimized clearing condition in the cardiac spheroid model. Results Here we first reported that the optical spheroid clearing protocol for the image-based toxicity prediction model. In our results, spheroid clearing significantly increases fluorescence intensity and enables image-based toxicity prediction. Conclusion We propose that these clearing methods of spheroid can be utilized for the image-based cardiotoxicity evaluation. Furthermore, we also present the possibility that our protocol can also be utilized for patient-tailored toxicity prediction evaluation.背景 基于二维细胞培养（two-dimensional cell culture）的毒性评估结果与临床结果存在差异，且无法准确反映细胞间的交叉信号传导，存在固有局限性。由于人体中几乎所有细胞均以三维结构（three-dimensional structure）排布并构成组织，因此由人体细胞构建的体外三维组织复制品，可作为药物开发与毒性评估的有效模型。组织透明化技术（clearing technique）可提升图像分辨率，实现全组织三维生物成像，从而可利用细胞球状体（spheroids）更高效地开展疾病模型分析与毒性评估。 方法 本研究制备了直径100~200微米的细胞球状体，并对透明化条件进行了优化。随后将优化后的透明化条件应用于心脏细胞球状体模型并完成成像。 结果 本研究首次报道了适用于基于图像的毒性预测模型的细胞球状体光学透明化方案。研究结果显示，细胞球状体透明化可显著提升荧光信号强度，为基于图像的毒性预测提供了可行支撑。 结论 本研究提出，上述细胞球状体透明化方法可应用于基于图像的心脏毒性（cardiotoxicity）评估。此外，本研究还证实该方案有望用于患者专属的毒性预测评估。