文献类型：期刊文献

英文题名：Efficient generation of biliary epithelial cells from rabbit intrahepatic bile duct by Y-27632 and Matrigel

作者：Jin, Lifang[1];Ji, Shaohui[1,3,4];Sun, Aijing[2]

机构：[1]Shaoxing Univ, Coll Life Sci, Shaoxing, Zhejiang, Peoples R China;[2]Zhejiang Univ, Shaoxing Hosp, Shaoxing Peoples Hosp, Dept Pathol, Shaoxing 312000, Zhejiang, Peoples R China;[3]Chinese Acad Sci, Kunming Primate Res Ctr, Kunming, Yunnan, Peoples R China;[4]Chinese Acad Sci, Kunming Inst Zool, Kunming, Yunnan, Peoples R China

年份：2013

卷号：49

期号：6

起止页码：433

外文期刊名：IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL

收录：SCI-EXPANDED(收录号：WOS:000320279500006)、、Scopus(收录号：2-s2.0-84879186540)、WOS

基金：This work was supported by research grants from Zhejiang Provincial Natural Science Foundation of China Y2110911 and Zhejiang Provincial Key Medical Discipline Construction Program of China GJSX-0100-004.

语种：英文

外文关键词：Rabbit; Biliary epithelial cells; Culture; Y-27632; Matrigel

外文摘要：Efficient culture of primary biliary epithelial cells (BECs) from adult liver is useful for both experimental studies and clinical applications of tissue engineering. However, an effective culture system for long-term proliferation of adult BECs is still unachieved. Laboratory rabbit has been used in a large number of studies; however, there are no reports of BECs from normal adult rabbit. As little as 5 g of normal rabbit liver tissue were minced, digested, and then clonally cultured in medium containing FBS and ITS. Cells were characterized by cell morphology, immunoassaying, and growth rate assay. Different combination of growth factors and substrates, including Y-27632 and Matrigel, were employed to assess their effect on cell proliferation. In the primary culture, the BECs cellular sheets consisting of cuboidal cells, as well as fibroblast-like cells and other hepatic cells, emerged with time of culture. The BECs cellular sheets were then manually split into cells clumps for further characterization. The subcultured cells had typical cell morphology of cholangiocytes, expressed the specific markers of BECs, including GGT, cytokeratin (CK18), and CK19, and possessed the capacity to form duct-like structure in three-dimensional Matrigel. Y-27632 and Matrigel-treated BECs had a steady growth rate as well as colony-formation capacity. The BECs were maintained in Y-27632 and Matrigel culture system for more than 3 mo. This is the first example, to our knowledge, of the successful culture of BECs from normal adult rabbit liver. Furthermore, our results indicate that treatment of BECs with Y-27632 and Matrigel is a simple method for efficient output of BECs.