title: Serum-free, long-term cultures of human hepatocytes: maintenance of cell morphology, transcription factors, and liver-specific functions.
abstract: Since human hepatocytes are available only in limited number, the development of a serum-free culture system for long-term cultivation of differentiated and functional hepatocytes is of great importance. Here we describe the culture of human hepatocytes in a chemically defined serum-free medium for up to 5 weeks. Cell morphology was assayed by light and electron microscopy and revealed a well-preserved cellular morphology. Marker proteins for epithelial and bile duct cells, cytokeratin (CK) 18 and 19, and liver-specific proteins, like phosphoenolpyruvate carboxykinase-2 (PCK2) and serum proteins, were expressed. Liver-enriched transcription factors CCAAT/enhancer binding protein alpha (C/EBPalpha) and hepatocyte nuclear factor-4 (HNF-4), cytokine and mitogen activated factors (nuclear factor kappa B) NFkappaB, and activator protein-1 (AP-1) were maintained and active for several weeks in our cultures. In summary, our serum-free culture system allows the culture of differentiated human hepatocytes for several weeks. It may serve as a model system for metabolic, pharmacologic-toxicologic studies, and studies on human pathogens under defined chemical conditions.
authors.affiliation: Klinik für Innere Medizin I, Martin Luther Universität Halle-Wittenberg, Halle, 06097, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Runge
authors.forename: D M
authors.initials: DM
authors.lastname: Runge
authors.forename: D
authors.initials: D
authors.lastname: Jäger
authors.forename: K A
authors.initials: KA
authors.lastname: Lubecki
authors.forename: D
authors.initials: D
authors.lastname: Beer Stolz
authors.forename: S
authors.initials: S
authors.lastname: Karathanasis
authors.forename: T
authors.initials: T
authors.lastname: Kietzmann
authors.forename: S C
authors.initials: SC
authors.lastname: Strom
authors.forename: K
authors.initials: K
authors.lastname: Jungermann
authors.forename: W E
authors.initials: WE
authors.lastname: Fleig
authors.forename: G K
authors.initials: GK
authors.lastname: Michalopoulos
mesh_terms: D001798:Blood Proteins / Q000235:genetics / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D015870:Gene Expression; D006801:Humans; D008099:Liver / Q000166:cytology* / Q000378:metabolism / Q000502:physiology*; D008854:Microscopy, Electron; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D001798:Blood Proteins; D016895:Culture Media, Serum-Free; D012333:RNA, Messenger; D014157:Transcription Factors