Isolation and characterization of bipotent liver progenitor cells from adult mouse.
Liver progenitor cells have drawn a great deal of attention both for their therapeutic potential and for their usefulness in exploring the molecular events surrounding liver development and regeneration. Despite the intensive studies on liver progenitors from rats, equivalent progenitor cells derived from mice are relatively rare. We used retrosine treatment followed by partial hepatectomy to elicit liver progenitors in mice. From these animals showing prominent ductular reactions, mouse-derived liver progenitor cell lines (LEPCs) were isolated by single-cell cloning. Phenotypic and lineage profiling of the LEPC clones were performed using immunochemistry, reverse transcription-polymerase chain reaction, and a dual-color system comprising the reporter EGFP under the control of the cytokeratin 19 promoter and the DsRed reporter under the control of the albumin promoter. LEPCs expressed liver progenitor cell markers. LEPCs also expressed some markers shared by bone marrow-derived hematopoietic stem cells c-Kit and Thy-1 but not CD34 and CD45. When cultured as aggregates in Matrigel, LEPCs differentiated into hepatocyte upon treatment with 50 ng/ml epithelial growth factor or differentiated into biliary lineage cells upon treatment with 20 ng/ml hepatocyte growth factor. In the presence of 2% dimethyl sulfoxide and 2% Matrigel, LEPCs acquired predominantly bile lineage phenotypes, with occasional patches of cells exhibiting hepatocyte phenotypes. Upon transplantation into CCl4-injured-liver, LEPCs engrafted into liver parenchyma and differentiated into hepatocytes. Considering the amenability of the mouse to genetic manipulation, these mouse-derived LEPCs may be useful tools as in vitro models to study molecular events in liver development and regeneration and can shed light in studying the therapy potential of liver stem cells.
Li
Wen-Lin
WL
Department of Cell Biology, Second Military Medical University, Xiangyin Road 800, Shanghai 200433, P.R. China.
Su
Juan
J
Yao
Yu-Cheng
YC
Tao
Xin-Rong
XR
Yan
Yong-Bi
YB
Yu
Hong-Yu
HY
Wang
Xin-Min
XM
Li
Jian-Xiu
JX
Yang
Yong-Ji
YJ
Lau
Joseph T Y
JT
Hu
Yi-Ping
YP
D000818:Animals; D001652:Bile Ducts / Q000166:cytology; D015415:Biomarkers / Q000032:analysis; D002454:Cell Differentiation; D049109:Cell Proliferation; D017690:Cell Transplantation; D004847:Epithelial Cells / Q000378:metabolism / Q000502:physiology; D006367:HeLa Cells; D006498:Hepatectomy; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D008099:Liver / Q000502:physiology* / Q000601:surgery; D008115:Liver Regeneration; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D041681:NIH 3T3 Cells; D011763:Pyrrolizidine Alkaloids / Q000494:pharmacology; D013234:Stem Cells / Q000378:metabolism* / Q000502:physiology; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection
D015415:Biomarkers; D011763:Pyrrolizidine Alkaloids; D014157:Transcription Factors; C003300:retrorsine