Mouse A6-positive hepatic oval cells derived from embryonic stem cells.
Oval cells have a potential to differentiate into a variety of cell lineages including hepatocytes and biliary epithelia. Several models have been established to activate the oval cells by incorporating a variety of toxins and carcinogens, alone or combined with surgical treatment. Those models are obviously not suitable for the study on human hepatic oval cells. It is necessary to establish a new and efficient model to study the human hepatic oval cells. In this study, the hepatocyte growth factor (HGF) and epidermal growth factor (EGF) were used to induce differentiation of mouse embryonic stem (ES) cells into hepatic oval cells. We first confirmed that hepatic oval cells derived from ES cells, which are bipotential, do exist during the course of mouse ES cells' differentiation into hepatic parenchymal cells. RT-PCR and transmission electron microscopy were applied in this study. The ratio of Sca-1+/CD34+ cells sorted by FACS in the induction group was increased from day 4 and reached the maximum on the day 8, whereas that in the control group remained at a low level. The differentiation ratio of Sca-1+/CD34+ cells in the induction group was significantly higher than that in the control group. About 92.48% of the sorted Sca-1+/CD34+ cells on the day 8 were A6 positive. Highly purified A6+/Sca-1+/CD34+ hepatic oval cells derived from ES cells could be obtained by FACS. The differentiation ratio of hepatic oval cells in the induction group (up to 4.46%) was significantly higher than that in the control group. The number of hepatic oval cells could be increased significantly by HGF and EGF. The study also examined the ultrastructures of ES-derived hepatic oval cells' membrane surface by atomic force microscopy. The ES-derived hepatic oval cells cultured and sorted by our protocols may be available for the future clinical application.
Department of General Surgery, Huangshi Central Hospital, Huangshi, 435000, China.|Department of General Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
Dong-Zhi
DZ
Yin
College of Life Science and Technology, Jinan University, Guangzhou, 510632, China. tjycai@jnu.edu.cn.
Ji-Ye
JY
Cai
Department of General Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
Qi-Chang
QC
Zheng
Department of Microbiology and Immunology, University of Illinois, College of Medicine, Chicago, IL, 60612, USA.
Zheng-Wei
ZW
Chen
College of Life Science and Technology, Jinan University, Guangzhou, 510632, China.
Jing-Xian
JX
Zhao
Department of General Surgery, Huangshi Central Hospital, Huangshi, 435000, China.
You-Neng
YN
Yuan
D000818:Animals; D018952:Antigens, CD34 / Q000235:genetics / Q000378:metabolism; D000950:Antigens, Ly / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005434:Flow Cytometry; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D017228:Hepatocyte Growth Factor / Q000494:pharmacology; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008840:Microfilament Proteins / Q000378:metabolism; D018625:Microscopy, Atomic Force; D046529:Microscopy, Electron, Transmission; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D013997:Time Factors
D018952:Antigens, CD34; D000950:Antigens, Ly; D004815:Epidermal Growth Factor; D017228:Hepatocyte Growth Factor; D008565:Membrane Proteins; D008840:Microfilament Proteins; C074650:Ly6a protein, mouse; C485190:Twf1 protein, mouse