Cholangiopathy is a diverse spectrum of chronic progressive bile duct disorders with limited treatment options and dismal outcomes. Scaffold- and stem cell-based tissue engineering technologies hold great promise for reconstructive surgery and tissue repair. Here, we report a combined application of 3D scaffold fabrication and reprogramming of patient-specific human hepatocytes to produce implantable artificial tissues that imitate the mechanical and biological properties of native bile ducts. The human chemically derived hepatic progenitor cells (hCdHs) were generated using two small molecules A83-01 and CHIR99021 and seeded inside the tubular scaffold engineered as a synergistic combination of two layers. The inner electrospun fibrous layer was made of nanoscale–macroscale polycaprolactone fibers acting to promote the hCdHs attachment and differentiation, while the outer microporous foam layer served to increase mechanical stability. The two layers of fiber and foam were fused robustly together thus creating coordinated mechanical flexibility to exclude any possible breaking during surgery. The gene expression profiling and histochemical assessment confirmed that hCdHs acquired the biliary epithelial phenotype and filled the entire surface of the fibrous matrix after 2 weeks of growth in the cholangiocyte differentiation medium in vitro. The fabricated construct replaced the macroscopic part of the common bile duct (CBD) and re-stored the bile flow in a rabbit model of acute CBD injury. Animals that received the acellular scaffolds did not survive after the replacement surgery. Thus, the artificial bile duct constructs populated with patient-specific hepatic progenitor cells could provide a scalable and compatible platform for treating bile duct diseases.

中文翻译：

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用化学重编程和微地形分化细胞进行生物工程的患者特异性胆管移植

胆管病是多种慢性进行性胆管疾病，具有有限的治疗选择和令人沮丧的结果。基于支架和干细胞的组织工程技术为重建手术和组织修复带来了巨大的希望。在这里，我们报告了 3D 支架制造和患者特异性人类肝细胞重新编程的组合应用，以生产模仿天然胆管的机械和生物学特性的可植入人造组织。使用两个小分子 A83-01 和 CHIR99021 生成人类化学衍生的肝祖细胞 (hCdHs)，并接种在管状支架内，该支架被设计为两层的协同组合。内部电纺纤维层由纳米级 - 宏观级聚己内酯纤维制成，用于促进 hCdHs 的附着和分化，而外部微孔泡沫层用于提高机械稳定性。两层纤维和泡沫牢固地融合在一起，从而产生协调的机械柔韧性，以排除手术过程中任何可能的断裂。基因表达谱和组织化学评估证实，在体外胆管细胞分化培养基中生长 2 周后，hCdHs 获得了胆管上皮表型并充满了纤维基质的整个表面。制造的结构取代了胆总管 (CBD) 的宏观部分，并在兔急性 CBD 损伤模型中恢复了胆汁流动。接受无细胞支架的动物在置换手术后无法存活。因此，填充有患者特异性肝祖细胞的人工胆管结构可以为治疗胆管疾病提供可扩展且兼容的平台。