Kuijk Ewart W, Rasmussen Shauna, Blokzijl Francis, Huch Meritxell, Gehart Helmuth, Toonen Pim, Begthel Harry, Clevers Hans, Geurts Aron M, Cuppen Edwin
Hubrecht Institute, KNAW and University Medical Center Utrecht, 3584CT Utrecht, The Netherlands.
University Medical Center Utrecht, PO Box 85060, 3508 AB Utrecht, The Netherlands.
Sci Rep. 2016 Feb 26;6:22154. doi: 10.1038/srep22154.
The rat is an important model for liver regeneration. However, there is no in vitro culture system that can capture the massive proliferation that can be observed after partial hepatectomy in rats. We here describe the generation of rat liver stem cell lines. Rat liver stem cells, which grow as cystic organoids, were characterized by high expression of the stem cell marker Lgr5, by the expression of liver progenitor and duct markers, and by low expression of hepatocyte markers, oval cell markers, and stellate cell markers. Prolonged cultures of rat liver organoids depended on high levels of WNT-signalling and the inhibition of BMP-signaling. Upon transplantation of clonal lines to a Fah(-/-) Il2rg(-/-) rat model of liver failure, the rat liver stem cells engrafted into the host liver where they differentiated into areas with FAH and Albumin positive hepatocytes. Rat liver stem cell lines hold potential as consistent reliable cell sources for pharmacological, toxicological or metabolic studies. In addition, rat liver stem cell lines may contribute to the development of regenerative medicine in liver disease. To our knowledge, the here described liver stem cell lines represent the first organoid culture system in the rat.
大鼠是肝脏再生的重要模型。然而，目前尚无体外培养系统能够模拟大鼠部分肝切除术后所观察到的大量细胞增殖现象。在此，我们描述了大鼠肝脏干细胞系的建立。以囊性类器官形式生长的大鼠肝脏干细胞，其特征在于干细胞标志物Lgr5的高表达、肝脏祖细胞和胆管标志物的表达以及肝细胞标志物、卵圆细胞标志物和星状细胞标志物的低表达。大鼠肝脏类器官的长期培养依赖于高水平的WNT信号传导和BMP信号传导的抑制。将克隆系移植到Fah(-/-) Il2rg(-/-) 大鼠肝衰竭模型后，大鼠肝脏干细胞植入宿主肝脏，并在其中分化为表达FAH和白蛋白的阳性肝细胞区域。大鼠肝脏干细胞系有望成为药理学、毒理学或代谢研究中稳定可靠的细胞来源。此外，大鼠肝脏干细胞系可能有助于肝脏疾病再生医学的发展。据我们所知，本文所述的肝脏干细胞系代表了大鼠首个类器官培养系统。
