Low asialoglycoprotein receptor expression as markers for highly proliferative potential hepatocytes.
Development of a reliable method to isolate highly proliferative potential hepatocytes will provide insight into the molecular mechanisms of liver regeneration, as well as proving crucial for the development of a biohybrid artificial liver. The aim of this study is to isolate highly proliferative, e.g., progenitor-like, hepatocytes. To this end, we fractionated hepatocytes expressing low and high levels of the asialoglycoprotein receptor (ASGP-R) based on the difference in their adhesion to poly[N-p-vinylbenzyl-O-beta-d-galactopyranosyl-(1-->4)-d-gluconamide] (PVLA), and examined the proliferative activity and gene expression of these fractionated hepatocytes. The results showed that approximately 0.5 to 1% of the total number of hepatocytes, which showed low adhesion to PVLA, expressed low levels of the ASGP-R, while the rest of hepatocyte population with high adhesion to PVLA expressed high levels of the ASGP-R. Interestingly hepatocytes with low ASGP-R expression levels had much higher DNA synthesizing activity (i.e., are much more proliferative) than those with high ASGP-R expression levels. Moreover, hepatocytes with low ASGP-R expression levels expressed higher levels of epidermal growth factor receptor (EGF-R), CD29 (beta1 integrin) and CD49f (alpha6 integrin) and lower levels of glutamine synthetase than those with high ASGP-R expression. These findings suggested that hepatocytes with low adhesion to PVLA due to their low ASGP-R expression could be potential candidates for progenitor-like hepatocytes due to their high proliferative capacity; hence, the low expression of the ASGP-R could be a unique marker for progenitor hepatocytes. The isolation of hepatocytes with different functional phenotypes using PVLA may provide a new research tool for a better understanding of the biology of hepatocytes and the mechanisms regulating their proliferation and differentiation in health and disease.
Ise
H
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Department of Organ Regeneration, Institute of Organ Transplants, Reconstructive Medicine and Tissue Engineering, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto 390-8621, Japan.
Sugihara
N
N
Negishi
N
N
Nikaido
T
T
Akaike
T
T
D000818:Animals; D037263:Asialoglycoprotein Receptor; D001212:Asialoglycoproteins / Q000502:physiology; D015415:Biomarkers / Q000032:analysis; D002448:Cell Adhesion / Q000502:physiology*; D002455:Cell Division / Q000502:physiology*; D002469:Cell Separation; D002478:Cells, Cultured; D066246:ErbB Receptors / Q000235:genetics / Q000502:physiology; D022781:Hepatocytes / Q000166:cytology* / Q000502:physiology*; D007785:Lactose / Q000031:analogs & derivatives; D008099:Liver / Q000166:cytology*; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D011137:Polystyrenes; D011956:Receptors, Cell Surface / Q000235:genetics* / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic
D037263:Asialoglycoprotein Receptor; D001212:Asialoglycoproteins; D015415:Biomarkers; D011137:Polystyrenes; D011956:Receptors, Cell Surface; C074112:poly-N-4-vinylbenzyllactonamide; D066246:ErbB Receptors; D007785:Lactose