Generation of insulin-secreting cells from mouse gallbladder stem cells by small molecules in vitro
Generation of insulin-secreting cells from mouse gallbladder stem cells by small molecules in vitro
复制标题
体外小分子诱导小鼠胆囊干细胞产生胰岛素分泌细胞
DOI：
10.1186/s13287-019-1407-6
复制
发表时间：
2019-09-23
影响因子：
7.5
通讯作者：
Wang, Min-Jun
中科院分区：
文献类型：
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作者：
Chen, Fei;Li, Tuo;Wang, Min-Jun
研究背景干细胞来源的胰岛β样细胞在治疗糖尿病方面前景广阔。胆囊属于肝外胆管系统，具有干细胞样细胞。这些干细胞可以在体外扩增，并具有分化为肝细胞、胆管细胞或胰腺细胞的潜力。由于胆囊具有很高的可利用性，因此，胆囊干细胞为胰岛β样细胞提供了新的细胞来源。方法以CK19Cre(ERT)；Rosa26R-GFP小鼠为研究对象，分离CK19(+)细胞，表达胆囊壁中的EpCAM(+)干细胞。在改良久保田培养液中培养扩增，并进行进一步分析。然后，我们开发了一种策略，以筛选能够从胆囊干细胞产生胰岛素分泌细胞的小分子组合。用胰腺细胞标记物鉴定这些细胞。最后将它们接种到纤维海绵中，移植到糖尿病小鼠体内进行功能检测。结果胆囊干细胞可以扩增15代以上。它们表达典型的肝干细胞标记物，包括CK19、EpCAM、Sox9和白蛋白。通过筛选方法，我们发现加入Noggin、FR180204和环多巴胺可以有效地诱导胆囊干细胞分化为胰岛素分泌细胞。这些细胞表达Pdx1、Nkx6.1和胰岛素，但不表达GCG。结论本研究提供了一种新的方法，可以在不经过基因修饰的情况下，从小鼠的胆囊中获得胰岛素分泌细胞。这为治疗1型糖尿病的β细胞疗法提供了一个选择。
BackgroundStem cell-derived pancreatic beta -like cells hold great promise for treating diabetes. Gallbladder belongs to the extrahepatic bile duct system and possesses stem-like cells. These stem cells could be expanded in vitro and have the potential of differentiating into hepatocytes, cholangiocytes, or pancreatic cells. As the gallbladder is highly available, gallbladder stem cells provide a new cell source of pancreatic beta -like cells. In this study, we aimed to investigate an approach for the generation of pancreatic beta -like cells from gallbladder stem cells (GSCs) without genetic modification.MethodsA CK19Cre(ERT);Rosa26R-GFP mouse was used to isolate CK19(+) cells, which represented EpCAM(+) stem cells in the gallbladder. They were cultured in the modified Kubota's medium for expansion and further analyzed. Then, we developed a strategy to screen a combination of small molecules that can generate insulin-secreting cells from gallbladder stem cells. These cells were identified with markers of pancreatic cells. Finally, they were seeded into the cellulosic sponge and transplanted to the diabetic mice for functional examination in vivo.ResultsGallbladder stem cells could be expanded for more than 15 passages. They expressed typical hepatic stem cell markers including CK19, EpCAM, Sox9, and albumin. By screening method, we found that adding Noggin, FR180204, and cyclopamine could efficiently induce gallbladder stem cells differentiating into insulin-secreting cells. These cells expressed Pdx1, Nkx6.1, and insulin but were negative for Gcg. After transplantation with the cellulosic sponge, they could ameliorate hyperglycemia in the diabetic mice.ConclusionThis study provides a new approach which can generate insulin-secreting cells from the gallbladder without genetic modification. This offers an option for beta cell therapy in treating type 1 diabetes.