Division of General and Thoracic Surgery, Translational Medicine Program, The Hospital for Sick Children, Toronto, ON M5G 1X8, Canada; Division of Pediatric Surgery, Department of Surgery, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
Division of General and Thoracic Surgery, Translational Medicine Program, The Hospital for Sick Children, Toronto, ON M5G 1X8, Canada.
J Pediatr Surg. 2021 Jan;56(1):11-16. doi: 10.1016/j.jpedsurg.2020.09.043. Epub 2020 Oct 6.
Biliary atresia (BA) is a fibro-obliterative cholangiopathy that involves both extrahepatic and intrahepatic bile ducts in infants. Cholangiocyte apoptosis has an influence on the fibrogenesis process of bile ducts and the progression of liver fibrosis in BA. Human amniotic fluid stem cells (hAFSCs) are multipotent cells that have ability to inhibit cell apoptosis. We aimed to investigate whether hAFSCs have the potential to attenuate cholangiocyte apoptosis and injury induced fibrogenic response in our ex vivo bile duct injury model of liver ductal organoids.
The anti-apoptotic effect of hAFSCs was tested in the acetaminophen-induced injury model of neonatal mouse liver ductal organoids (AUP #42681) by using direct and indirect co-culture systems. Cell apoptosis and proliferation were evaluated by immunofluorescent staining. Expression of fibrogenic cytokines was analyzed by RT-qPCR. Data were compared using one-way ANOVA with post hoc test.
In our injury model, liver ductal organoids that were treated with hAFSCs in both direct and indirect co-culture systems had a significantly smaller number of apoptotic cholangiocytes and decreased expression of fibrogenic cytokines, transforming growth factor beta-1 (TGF-β1) and platelet-derived growth factor-BB (PDGF-BB). Moreover, hAFSCs increased cholangiocyte proliferation in injured organoids.
hAFSCs have the ability to protect the organoids from injury by decreasing cholangiocyte apoptosis and promoting cholangiocyte proliferation. This protective ability of hAFSCs leads to inhibition of the fibrogenic response in the injured organoids. hAFSCs have high therapeutic potential to attenuate liver fibrogenesis in cholangiopathic diseases such as BA.
胆道闭锁（BA）是一种纤维性阻塞性胆管病，涉及婴儿的肝外和肝内胆管。胆管细胞凋亡对胆管纤维化过程和 BA 中肝纤维化的进展有影响。人羊水干细胞（hAFSCs）是多能细胞，具有抑制细胞凋亡的能力。我们旨在研究 hAFSCs 是否有可能在我们的胆管上皮细胞类器官肝内胆管损伤的体外模型中减轻胆管细胞凋亡和损伤诱导的纤维生成反应。
通过直接和间接共培养系统，在乙酰氨基酚诱导的新生小鼠肝内胆管上皮细胞类器官（AUP#42681）损伤模型中测试 hAFSCs 的抗凋亡作用。通过免疫荧光染色评估细胞凋亡和增殖。通过 RT-qPCR 分析纤维生成细胞因子的表达。使用单因素方差分析和事后检验比较数据。
在我们的损伤模型中，直接和间接共培养系统中 hAFSCs 处理的肝内胆管上皮细胞类器官，其凋亡的胆管细胞数量明显减少，纤维生成细胞因子转化生长因子-β1（TGF-β1）和血小板衍生生长因子-BB（PDGF-BB）的表达降低。此外，hAFSCs 增加了受损类器官中胆管细胞的增殖。
hAFSCs 具有通过减少胆管细胞凋亡和促进胆管细胞增殖来保护类器官免受损伤的能力。hAFSCs 的这种保护能力导致受损类器官中纤维生成反应的抑制。hAFSCs 具有减轻胆道疾病如 BA 中肝纤维化的高治疗潜力。
