title: Morphological changes induced by extracellular matrix are correlated with maturation of rat small hepatocytes.
abstract: Small hepatocytes (SHs), which are known to be hepatic progenitor cells, were isolated from an adult rat liver. SHs in a colony sometimes change their shape from small to large and from flat to rising/piled-up. The aim of the present study is to clarify whether the alteration of cell shape is correlated with the maturation of SHs and whether extracellular matrix (ECM) can induce the morphological changes of SHs. We used liver-enriched transcription factors (LETFs) such as hepatocyte nuclear factor (HNF) 4 alpha, HNF6, CCAAT/enhancer binding proteins (C/EBP) alpha, and C/EBP beta, tryptophan 2,3-dioxygenase (TO), and serine dehydratase (SDH) as markers of hepatic maturation. To enrich the number of SH colonies, the colonies were isolated from dishes and replated. Replated colonies proliferated and the average number of cells per colony was about five times larger at day 9 than at day 1. When the cells were treated with laminin, type IV collagen, a mixture of laminin and type IV collagen, Matrigel or collagen gel (CG), only the cells treated with Matrigel dramatically changed their shape within several days and had reduced growth activity, whereas the cells treated with other ECM did not. HNF4 alpha, HNF6, C/EBP alpha, C/EBP beta, and TO were well expressed in the cells treated with Matrigel. Furthermore, addition of both glucagon and dexamethasone dramatically induced the expression of SDH mRNA and protein in the cells treated with Matrigel. In conclusion, morphological changes of SHs may be correlated with hepatic maturation and basement membrane (BM)-like structure may induce the morphological changes of SHs.
authors.affiliation: Department of Pathology, Cancer Research Institute, Sapporo Medical University School of Medicine, Chuo-Ku, S-1, W-17, Sapporo 060-8556, Japan.
authors.forename: Shinichi
authors.initials: S
authors.lastname: Sugimoto
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Mitaka
authors.forename: Shinichiro
authors.initials: S
authors.lastname: Ikeda
authors.forename: Keisuke
authors.initials: K
authors.lastname: Harada
authors.forename: Iwao
authors.initials: I
authors.lastname: Ikai
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Yamaoka
authors.forename: Yohichi
authors.initials: Y
authors.lastname: Mochizuki
mesh_terms: D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D022763:CCAAT-Enhancer-Binding Protein-alpha / Q000378:metabolism; D022782:CCAAT-Enhancer-Binding Protein-beta / Q000378:metabolism; D002455:Cell Division; D002478:Cells, Cultured; D003094:Collagen / Q000494:pharmacology; D004268:DNA-Binding Proteins; D004338:Drug Combinations; D004797:Enzyme-Linked Immunosorbent Assay; D005109:Extracellular Matrix / Q000378:metabolism*; D051557:Hepatocyte Nuclear Factor 4; D051559:Hepatocyte Nuclear Factor 6; D022781:Hepatocytes / Q000166:cytology*; D018398:Homeodomain Proteins / Q000378:metabolism; D007150:Immunohistochemistry; D012695:L-Serine Dehydratase / Q000378:metabolism; D007797:Laminin / Q000378:metabolism / Q000494:pharmacology; D008099:Liver / Q000378:metabolism*; D008858:Microscopy, Phase-Contrast; D010750:Phosphoproteins / Q000378:metabolism; D011509:Proteoglycans / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013234:Stem Cells / Q000378:metabolism; D013997:Time Factors; D015534:Trans-Activators / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D014366:Tryptophan Oxygenase / Q000378:metabolism
chemical_list: D022763:CCAAT-Enhancer-Binding Protein-alpha; D022782:CCAAT-Enhancer-Binding Protein-beta; D004268:DNA-Binding Proteins; D004338:Drug Combinations; D051557:Hepatocyte Nuclear Factor 4; D051559:Hepatocyte Nuclear Factor 6; C496594:Hnf4a protein, rat; D018398:Homeodomain Proteins; D007797:Laminin; C496607:Onecut1 protein, rat; D010750:Phosphoproteins; D011509:Proteoglycans; D015534:Trans-Activators; D014157:Transcription Factors; C059485:matrigel; D003094:Collagen; D014366:Tryptophan Oxygenase; D012695:L-Serine Dehydratase