肝器官样品已成为研究肝脏生物学和疾病、开发新疗法和再生医学方法的有力工具。对于器官样品培养，Matrigel是最常用的一种细胞外基质。然而，由于存在未知蛋白质引起的免疫排斥、批次间变异和血管生成问题，Matrigel不能用于临床应用。为了获得人类原代肝细胞(hPHs)，我们进行了两步胶原酶肝灌注方案。我们使用三种小分子药物（A83-01，CHIR99021和HGF）来重编程hPHs为人类化学来源的肝前体细胞（hCdHs），并使用hCdHs来生成肝器官样品。 在本研究中，我们报告了使用hCdHs在胶原支架上生成肝器官样品。与胶原支架上成人肝脏（或原代肝细胞）样品（hALO_C）相比，hCdHs派生的胶原支架上肝器官样品（hCdHO_C）显示出10倍的器官样品生成效率增加，并具有更高的肝脏或肝前体特异性标记的表达。此外，我们证明了hCdHO_C能够分化为肝脏样品（hCdHO_C_DM），表明这些器官样品具有作为药物筛选平台的潜力。 总体而言，我们的研究突出了hCdHO_C作为肝脏研究工具的潜力，并提出了使用hCdHs和胶原支架生成肝器官样品的新方法。

Liver organoids have emerged as a powerful tool for studying liver biology and disease and for developing new therapies and regenerative medicine approaches. For organoid culture, Matrigel, a type of extracellular matrix, is the most commonly used material. However, Matrigel cannot be used for clinical applications due to the presence of unknown proteins that can cause immune rejection, batch-to-batch variability, and angiogenesis.To obtain human primary hepatocytes (hPHs), we performed 2 steps collagenase liver perfusion protocol. We treated three small molecules cocktails (A83-01, CHIR99021, and HGF) for reprogramming the hPHs into human chemically derived hepatic progenitors (hCdHs) and used hCdHs to generate liver organoids.In this study, we report the generation of liver organoids in a collagen scaffold using hCdHs. In comparison with adult liver (or primary hepatocyte)-derived organoids with collagen scaffold (hALO_C), hCdH-derived organoids in a collagen scaffold (hCdHO_C) showed a 10-fold increase in organoid generation efficiency with higher expression of liver- or liver progenitor-specific markers. Moreover, we demonstrated that hCdHO_C could differentiate into hepatic organoids (hCdHO_C_DM), indicating the potential of these organoids as a platform for drug screening.Overall, our study highlights the potential of hCdHO_C as a tool for liver research and presents a new approach for generating liver organoids using hCdHs with a collagen scaffold.