用于毒理学和功能研究的患者来源肝脏类器官生物样本库的开发与表征

Liver is the largest solid internal organ and performs numerous critical functions. Annually, liver diseases claim about 2 million lives globally (1). Traditional models for investigating liver biology, including 2D cultured cell lines, primary cells, and 3D spheroids, fail to accurately represent the in vivo metabolic and structural complexities. Additionally, animal models are expensive, time-consuming, ineffective, and may pose ethical dilemmas. Therefore, there is an urgent need for advanced liver models that can accurately mimic tissue characteristics.Organoids are self-organizing 3D structures that replicate the architecture and functionality of the original tissue (2). Patient-derived organoids (PDOs) closely resemble the source tissue and may reflect individual responses to therapy. In this study, we present the development and characterization of a biobank consisting of 11 individual liver PDO lines, which capture donor variability and enhance the applicability of these models for toxicology studies.Liver tissues were collected with donor consent. PDOs were created by adapting a previously published protocol (3). Briefly, tissue samples were minced, digested, filtered, combined with Matrigel (Corning), and cultured in liver PDO-specific growth media. When suitable, undifferentiated PDOs were matured using tailored media.PDO lines were effectively expanded and cryopreserved, demonstrating their robustness. All lines expressed liver-specific biomarkers, verified by qPCR and confocal microscopy. Each line was confirmed to be free of infectious agents and validated as unique via short tandem repeat analysis. Upon maturation, these lines exhibited increased albumin and urea production, along with inducible cytochrome P450 (CYP) activity, confirming their functional maturity. Donor-dependent variability in these phenotypes indicated that the biobank may represent the patient population. Four liver PDO lines were also evaluated for their response to 3 DILI drugs. Based on these results, the liver biobank was concluded to be a robust in vitro model for testing drug responses.Our liver PDO biobank reflects the patient population and will be a valuable tool for pharmaceutical clients. We have successfully established and characterized 11 liver PDO lines, with further characterization efforts ongoing. These PDOs will significantly advance our understanding of liver biology and disease mechanisms, offering applications in drug testing, including DMPK and ADME/ Tox studies. Future initiatives will focus on scaling up, developing specific assays, and creating PDOs from patients with liver conditions, such as hepatocellular carcinoma. References: 1.Asrani et al., 2019. J. Hepatol.2.Zhao et al., 2022. STAR Protocols3.Broutier et al., 2016. Nature Protocols