Peribiliary Glands Are Key in Regeneration of the Human Biliary Epithelium After Severe Bile Duct Injury
Peribiliary Glands Are Key in Regeneration of the Human Biliary Epithelium After Severe Bile Duct Injury
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DOI：
10.1002/hep.30365
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发表时间：
2019-04-01
期刊：
影响因子：
13.5
通讯作者：
Porte, Robert J.
中科院分区：
文献类型：
--
作者：
de Jong, Iris E. M.;Matton, Alix P. M.;Porte, Robert J.
胆管周围腺（PBG）是组织在环绕大胆管的细胞网络中的干细胞/祖细胞的来源。已提出伴有管腔胆管上皮丧失的严重胆管病可激活PBG，导致细胞增殖和分化以恢复胆管上皮完整性。然而，在人类肝脏中缺乏这一概念的正式证据。因此，我们开发了一种离体模型，使用从丢弃的供体肝脏中获得的肝外人胆管的精确切割切片，提供了细胞结构的完整解剖组织，以研究严重胆道损伤后PBG细胞的时空分化和迁移。将缺血后胆管切片在含氧培养基中孵育长达一周。在基线时，严重的组织损伤是明显的，管腔上皮衬里和壁间质坏死的损失。相反，PBG保持相对较好的保存和不同的反应，PBG注意到，包括PBG扩张，细胞增殖和成熟。氧合孵育24小时后，PBG细胞增殖增加，72小时后达到高峰。PBG细胞的增殖通过PBG细胞凋亡减少和从原始和多能（同源异型盒蛋白Nanog+/性别决定区Y-box 9+）分化为成熟（囊性纤维化跨膜传导调节因子+/分泌素受体+）和活化表型（缺氧诱导因子1 α、葡萄糖转运蛋白1和血管内皮生长因子A表达增加）来抑制。在我们的离体模型中，增殖的PBG细胞的迁移是无组织的，但导致在基质表面产生上皮单层。结论：人PBG含有胆管祖细胞，并且能够通过增殖、分化和成熟来响应胆管上皮损失以恢复上皮完整性。人PBG细胞的离体时空行为为PBG在严重损伤后的胆管再生中的关键作用提供了证据。
Peribiliary glands (PBG) are a source of stem/progenitor cells organized in a cellular network encircling large bile ducts. Severe cholangiopathy with loss of luminal biliary epithelium has been proposed to activate PBG, resulting in cell proliferation and differentiation to restore biliary epithelial integrity. However, formal evidence for this concept in human livers is lacking. We therefore developed an ex vivo model using precision-cut slices of extrahepatic human bile ducts obtained from discarded donor livers, providing an intact anatomical organization of cell structures, to study spatiotemporal differentiation and migration of PBG cells after severe biliary injury. Postischemic bile duct slices were incubated in oxygenated culture medium for up to a week. At baseline, severe tissue injury was evident with loss of luminal epithelial lining and mural stroma necrosis. In contrast, PBG remained relatively well preserved and different reactions of PBG were noted, including PBG dilatation, cell proliferation, and maturation. Proliferation of PBG cells increased after 24 hours of oxygenated incubation, reaching a peak after 72 hours. Proliferation of PBG cells was paralleled by a reduction in PBG apoptosis and differentiation from a primitive and pluripotent (homeobox protein Nanog+/ sex-determining region Y-box 9+) to a mature (cystic fibrosis transmembrane conductance regulator+/secretin receptor+) and activated phenotype (increased expression of hypoxia-inducible factor 1 alpha, glucose transporter 1, and vascular endothelial growth factor A). Migration of proliferating PBG cells in our ex vivo model was unorganized, but resulted in generation of epithelial monolayers at stromal surfaces. Conclusion: Human PBG contain biliary progenitor cells and are able to respond to bile duct epithelial loss with proliferation, differentiation, and maturation to restore epithelial integrity. The ex vivo spatiotemporal behavior of human PBG cells provides evidence for a pivotal role of PBG in biliary regeneration after severe injury.