胆管细胞类器官为从体外建模到再生医学的组织工程的应用提供了强大的平台。然而,它们的扩增和分化通常在动物衍生的水凝胶中进行,阻碍类器官完全成熟为功能性胆管细胞。此外,这些水凝胶定义不清和复杂,限制类器官的临床适用性。在这项研究中,一种新的培养基组合物与合成聚异氰酸肽(PIC)水凝胶结合,以增强肝内胆管细胞类器官(ICO)成熟为功能性胆管细胞。在丁酸钠和丙戊酸存在下培养的ICO,组蛋白脱乙酰酶抑制剂,和一个Notch信号激活器,分别,在PIC水凝胶中表现出更成熟的表型,关键胆管细胞标志物的表达增加证明了这一点,对胆道功能至关重要。值得注意的是,PIC水凝胶中成熟的胆管细胞类器官显示顶端极性,与在Matrigel中培养的ICO的传统基底外极化相反。此外,这些成熟的胆管细胞类器官有效地模拟了转化生长因子β诱导的胆道促纤维化反应。一起来看,无动物,化学定义的培养系统,可促进ICO进入具有顶端极性的成熟胆管细胞,促进需要进入根尖膜的再生医学应用和体外研究,已开发。
Cholangiocyte organoids provide a powerful platform for applications ranging from in vitro modeling to tissue engineering for regenerative medicine. However, their expansion and differentiation are typically conducted in animal-derived hydrogels, which impede the full maturation of organoids into functional cholangiocytes. In addition, these hydrogels are poorly defined and complex, limiting the clinical applicability of organoids. In this study, a novel medium composition combined with synthetic polyisocyanopeptide (PIC) hydrogels to enhance the maturation of intrahepatic cholangiocyte organoids (ICOs) into functional cholangiocytes is utilized. ICOs cultured in the presence of sodium butyrate and valproic acid, a histone deacetylase inhibitor, and a Notch signaling activator, respectively, in PIC hydrogel exhibit a more mature phenotype, as evidenced by increased expression of key cholangiocyte markers, crucial for biliary function. Notably, mature cholangiocyte organoids in PIC hydrogel display apical-out
polarity, in contrast to the traditional basal-out polarization of ICOs cultured in Matrigel. Moreover, these mature cholangiocyte organoids effectively model the biliary pro-fibrotic response induced by transforming growth factor beta. Taken together, an animal-free, chemically defined culture system that promotes the ICOs into mature cholangiocytes with apical-out
polarity, facilitating regenerative medicine applications and in vitro studies that require access to the apical membrane, is developed.