Li Bin, Dorrell Craig, Canady Pamela S, Wakefield Leslie
Oregon Stem Cell Center, Oregon Health and Science University, Portland, OR, USA.
Methods Mol Biol. 2019;1905:19-27. doi: 10.1007/978-1-4939-8961-4_3.
Cholangiocytes are proliferative and are one of the sources for liver progenitor cells. Clonogenic cholangiocytes are defined as cells capable of clonally proliferating and differentiating cholangiocytes both in vitro and in vivo. In this protocol, we describe the method for isolation of primary cholangiocytes from mouse. To study the heterogeneity of cholangiocytes, we used flow cytometry-based cell sorting to isolate different subsets of cholangiocytes. Organoid-forming efficiencies from sorted single cells are compared within different cholangiocyte populations to identify clonogenic cholangiocytes.
胆管细胞具有增殖能力，是肝祖细胞的来源之一。克隆性胆管细胞被定义为能够在体外和体内进行克隆增殖并分化为胆管细胞的细胞。在本方案中，我们描述了从小鼠中分离原代胆管细胞的方法。为了研究胆管细胞的异质性，我们使用基于流式细胞术的细胞分选来分离胆管细胞的不同亚群。在不同的胆管细胞群体中比较分选后的单细胞形成类器官的效率，以鉴定克隆性胆管细胞。
