Willemse Jorke, van Tienderen Gilles, van Hengel Eline, Schurink Ivo, van der Ven Diana, Kan Yik, de Ruiter Petra, Rosmark Oskar, Westergren-Thorsson G Gunilla, Schneeberger Kerstin, van der Eerden Bram, Roest Henk, Spee Bart, van der Laan Luc, de Jonge Jeroen, Verstegen Monique
Department of Surgery, Transplant Institute, Erasmus MC, University Medical Center Rotterdam, the Netherlands.
Lung Biology, Department Experimental Medical Science, Lund University, Lund, Sweden.
Biomaterials. 2022 May;284:121473. doi: 10.1016/j.biomaterials.2022.121473. Epub 2022 Mar 24.
Human cholangiocyte organoids are promising for regenerative medicine applications, such as repair of damaged bile ducts. However, organoids are typically cultured in mouse tumor-derived basement membrane extracts (BME), which is poorly defined, highly variable and limits the direct clinical applications of organoids in patients. Extracellular matrix (ECM)-derived hydrogels prepared from decellularized human or porcine livers are attractive alternative culture substrates. Here, the culture and expansion of human cholangiocyte organoids in liver ECM(LECM)-derived hydrogels is described. These hydrogels support proliferation of cholangiocyte organoids and maintain the cholangiocyte-like phenotype. The use of LECM hydrogels does not significantly alter the expression of selected genes or proteins, such as the cholangiocyte marker cytokeratin-7, and no species-specific effect is found between human or porcine LECM hydrogels. Proliferation rates of organoids cultured in LECM hydrogels are lower, but the differentiation capacity of the cholangiocyte organoids towards hepatocyte-like cells is not altered by the presence of tissue-specific ECM components. Moreover, human LECM extracts support the expansion of ICO in a dynamic culture set up without the need for laborious static culture of organoids in hydrogel domes. Liver ECM hydrogels can successfully replace tumor-derived BME and can potentially unlock the full clinical potential of human cholangiocyte organoids.
人胆管细胞类器官在再生医学应用中具有广阔前景，比如用于修复受损胆管。然而，类器官通常在源自小鼠肿瘤的基底膜提取物（BME）中培养，这种提取物成分不明确、高度可变，限制了类器官在患者中的直接临床应用。由脱细胞人肝脏或猪肝脏制备的细胞外基质（ECM）衍生水凝胶是颇具吸引力的替代培养底物。在此，描述了人胆管细胞类器官在肝脏ECM（LECM）衍生水凝胶中的培养与扩增。这些水凝胶支持胆管细胞类器官的增殖并维持胆管细胞样表型。使用LECM水凝胶不会显著改变所选基因或蛋白质的表达，如胆管细胞标志物细胞角蛋白-7，并且在人或猪LECM水凝胶之间未发现物种特异性效应。在LECM水凝胶中培养的类器官增殖率较低，但胆管细胞类器官向肝细胞样细胞的分化能力不会因组织特异性ECM成分的存在而改变。此外，人LECM提取物在动态培养设置中支持胆管内凹（ICO）的扩增，而无需在水凝胶穹顶中对类器官进行费力的静态培养。肝脏ECM水凝胶能够成功替代肿瘤衍生的BME，并有可能释放人胆管细胞类器官的全部临床潜力。
