Tubular bile duct structure mimicking bile duct morphogenesis for prospective in vitro liver metabolite recovery
Tubular bile duct structure mimicking bile duct morphogenesis for prospective in vitro liver metabolite recovery
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DOI：
10.1186/s13036-020-0230-z
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发表时间：
2020-03-19
影响因子：
5.6
通讯作者：
Sakai, Yasuyuki
中科院分区：
文献类型：
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作者：
Safitri, Astia;Shinohara, Marie;Sakai, Yasuyuki
背景在临床药代动力学中，肝脏代谢产物被用于疾病诊断和药物检查。因此，建立一套重现肝脏代谢物回收的体外检测系统将为药学研究提供重要的益处。然而，肝脏模型的发展已被证明具有挑战性，因为缺乏合适的胆管结构来积累和运输来自肝实质的代谢物。目前可用的胆管模型，如胆管囊腔包埋的细胞外基质(ECM)，在形态上与活体胆管的管状形态缺乏任何相似之处。此外，这些系统不能克服代谢物回收问题，因为它们建立在孤立的培养系统中。在这里，我们成功地在开放培养系统中建立了一个与活体结构非常相似的非连续管状胆管结构模型。利用该系统可以有效地从肝实质细胞中分离出肝脏代谢物。结果原代大鼠肝母细胞、大鼠胆管上皮细胞和小鼠胚胎成纤维细胞三种细胞共培养可模拟肝脏发育过程中胆管的形态发生。用含有Matrigel和I型胶原凝胶混合物的ECM覆盖细胞，促进了管状胆管结构的发展。在该培养体系中，与胆管上皮细胞分化相关的特异性标志物和信号分子的表达在胆管形成过程中高度上调。这种胆管结构也使代谢物类似物从肝实质细胞中分离积累。结论一种基于形态发生的培养系统有效地建立了先进的胆管结构，提高了适合于自体体体外代谢物-胆汁采集的肝脏模型的可塑性，这可能会提高高通量肝模型在基于细胞的检测中的性能。
Background Liver metabolites are used to diagnose disease and examine drugs in clinical pharmacokinetics. Therefore, development of an in vitro assay system that reproduces liver metabolite recovery would provide important benefits to pharmaceutical research. However, liver models have proven challenging to develop because of the lack of an appropriate bile duct structure for the accumulation and transport of metabolites from the liver parenchyma. Currently available bile duct models, such as the bile duct cyst-embedded extracellular matrix (ECM), lack any morphological resemblance to the tubular morphology of the living bile duct. Moreover, these systems cannot overcome metabolite recovery issues because they are established in isolated culture systems. Here, we successfully established a non-continuous tubular bile duct structure model in an open-culture system, which closely resembled an in vivo structure. This system was utilized to effectively collect liver metabolites separately from liver parenchymal cells. Results Triple-cell co-culture of primary rat hepatoblasts, rat biliary epithelial cells, and mouse embryonic fibroblasts was grown to mimic the morphogenesis of the bile duct during liver development. Overlaying the cells with ECM containing a Matrigel and collagen type I gel mixture promoted the development of a tubular bile duct structure. In this culture system, the expression of specific markers and signaling molecules related to biliary epithelial cell differentiation was highly upregulated during the ductal formation process. This bile duct structure also enabled the separate accumulation of metabolite analogs from liver parenchymal cells. Conclusions A morphogenesis-based culture system effectively establishes an advanced bile duct structure and improves the plasticity of liver models feasible for autologous in vitro metabolite-bile collection, which may enhance the performance of high-throughput liver models in cell-based assays.