Development of Bifunctional Three-Dimensional Cysts from Chemically Induced Liver Progenitors
Development of Bifunctional Three-Dimensional Cysts from Chemically Induced Liver Progenitors
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DOI：
10.1155/2019/3975689
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发表时间：
2019-09-03
影响因子：
4.3
通讯作者：
Eguchi, Susumu
中科院分区：
文献类型：
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作者：
Huang, Yu;Sakai, Yusuke;Eguchi, Susumu
化学诱导的肝祖细胞（CLiP）在肝脏再生医学中具有广阔的应用前景。由肝祖细胞生成的三维（3D）结构在细胞移植、疾病模型和药物测试中具有广泛的应用。在这里，我们报告了 3D 囊状结构的自发形成，该结构包括在明胶涂层培养皿上成熟的大鼠 CLiP。我们的 3D 囊肿含有 Alb(+/+)CK19(+/-) 和 Ck19(+/+)Alb(+/-) 细胞。这些细胞类型逐渐分化为专门的成熟细胞，成熟胆道标志物（Cftr、Ae2 和 Aqp1）和肝脏标志物（Alb 和 Mrp2）的表达证明了这一点。 3D 囊肿还表达功能性多药耐药蛋白 1 (Mdr1)，如罗丹明上皮流出所示。此外，我们观察到肝细胞和胆酰-赖氨酰-荧光素挤出物之间的胆小管功能，表明 3D 囊肿和活性胆盐输出泵 (Bsep) 转运蛋白的功能特征完好无损。因此，我们的研究揭示了大鼠 CLiPs 自发形成 3D 囊状结构并伴随细胞体外成熟的自然特征，为肝脏发育和药物筛选的研究提供了平台。
Chemically induced liver progenitors (CLiPs) have promising applications in liver regenerative medicine. Three-dimensional (3D) structures generated from liver progenitor cells possess wide applications in cell transplantation, disease model, and drug testing. Here, we report on the spontaneous formation of 3D cystic structures comprising maturing rat CLiPs on gelatin-coated dishes. Our 3D cysts contained Alb(+/+)CK19(+/-) and Ck19(+/+)Alb(+/-) cells. These cell types gradually diverged into specialized mature cells, as demonstrated by the expression of mature biliary markers (Cftr, Ae2, and Aqp1) and hepatic markers (Alb and Mrp2). The 3D cysts also expressed functional multidrug resistance protein 1 (Mdr1), as indicated by epithelial efflux of rhodamine. Furthermore, we observed bile canaliculi functions between hepatocytes and cholyl-lysyl-fluorescein extrusions, indicating that the functional characteristics of 3D cysts and active bile salt export pump (Bsep) transporters were intact. Thus, our study revealed a natural characteristic of rat CLiPs to spontaneously form 3D cystic structures accompanied with cell maturation in vitro, offering a platform for studies of liver development and drug screening.