title: Maintenance of hepatic differentiation by hepatocyte attachment peptides derived from laminin chains.
abstract: Hepatocytes rapidly lose hepatic functions upon isolation from liver, perhaps due to disrupted cell/matrix interactions. The matrix macromolecule laminin-111 consists of three chains, α1, β1, and γ1; it is a major component of Matrigel, which can maintain hepatic differentiation. We previously showed that the A13 peptide (RQVFQVAYIIIKA, α1 chain 121-133) derived from mouse laminin α1 exhibits hepatocyte attachment activity and maintains hepatic differentiation. Here, we sought to identify hepatocyte adhesive sequences from the mouse laminin β1 and γ1 chains using 22 synthetic peptides that show biological activity for fibrosarcoma cells. Nine peptides showed hepatocyte attachment activity. Of these, B160 (VILQQSAADIAR, β1 chain 1607-1618), and C16 (KAFDITYVRLKF γ1 chain 139-150) exhibited the most potent activity. Hepatocytes cultured on both peptides also maintained expression of albumin, tyrosine aminotransferase, tryptophan-2,3-dioxygenase, and cytochrome P450. The morphology of hepatocytes on both peptides was a rounded shape typical for hepatic differentiation. We also characterized the nature of adhesion to the peptides. Heparin and EDTA inhibited cell attachment to both peptides, suggesting that hepatocyte attachment to the peptides was mediated by multiple receptors. The identification of active sequences regulating hepatic functions may facilitate the design of hepatocyte culture substrata that can regulate specific cellular behaviors in the context of a bioartificial liver.
authors: Laboratory of Clinical Biochemistry, Tokyo University of Pharmacy and Life Sciences, Tokyo, Japan. kikkawa@toyaku.ac.jp.
authors: Yamato
authors: Y
authors: Kikkawa
authors: Akira
authors: A
authors: Kataoka
authors: Yuji
authors: Y
authors: Matsuda
authors: Naoya
authors: N
authors: Takahashi
authors: Takahiro
authors: T
authors: Miwa
authors: Fumihiko
authors: F
authors: Katagiri
authors: Kentaro
authors: K
authors: Hozumi
authors: Motoyoshi
authors: M
authors: Nomizu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D015415:Biomarkers / Q000378:metabolism; D002448:Cell Adhesion; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D022781:Hepatocytes / Q000166:cytology / Q000502:physiology*; D007797:Laminin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D015415:Biomarkers; D007797:Laminin; D010455:Peptides; D021122:Protein Subunits