Generation of a monoclonal antibody specifically reacting with undifferentiated liver progenitor cells.
An adult rat liver progenitor cell line Lig-8 was established. In the induction by sodium butyrate, these cells were shown to be able to differentiate into both hepatocytes and bile duct cells expressing albumin and cytokeratin-19, the markers of respective cell types. In order to generate Lig-8 specific antibody for further studies, we produced a monoclonal antibody using the whole Lig-8 cells as immunogen. The yielded monoclonal antibody, named Ligab, belongs to IgG subclass G1 and kappa light chain. It specifically stained on Lig-8 cells in the cytoplasm but not on a rat hepatoma cell line H4IIE. Its immunoreaction against Lig-8 cell lysate on dot blots diminished as the concentration of sodium dodecyl sulfate (SDS) in the lysate increased to 2%, a level in the sample buffer of standard SDS-polyacrylamide gel electrophoresis (PAGE). Not surprisingly, Ligab failed to detect its reacting antigen in Lig-8 cell lysate by standard SDS-PAGE-based immunoblotting. It could detect this antigen only by native PAGE-based immunoblotting. These characteristics suggested that the antigenic epitope for Ligab is likely a molecular structure instead of a peptide sequence. More interestingly, expression of Ligab-reacting antigen in Lig-8 cells declined as the cells were induced to differentiate by sodium butyrate. This antigen is very likely a differentiation-related marker for these cells, and this monoclonal antibody may help study the molecular mechanisms of liver progenitor cell differentiation.
Chiu
Chien-Chang
CC
The Graduate Institute of Applied Science and Engineering, Fu-Jen Catholic University, Taipei County, Taiwan.
Huang
Yu-Wen
YW
Chou
Shiu-Huey
SH
Huang
Guan-Tarn
GT
Chiou
Ling-Ling
LL
Lee
Hsuan-Shu
HS
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology*; D002087:Butyrates; D002454:Cell Differentiation / Q000276:immunology; D045744:Cell Line, Tumor; D004591:Electrophoresis, Polyacrylamide Gel; D006825:Hybridomas / Q000276:immunology*; D015151:Immunoblotting; D008099:Liver / Q000166:cytology*; D008856:Microscopy, Fluorescence; D051381:Rats; D013234:Stem Cells / Q000276:immunology*
D000911:Antibodies, Monoclonal; D002087:Butyrates