title: Differentiation of human embryonic stem cells to hepatocytes using deleted variant of HGF and poly-amino-urethane-coated nonwoven polytetrafluoroethylene fabric.
abstract: Human embryonic stem (hES) cells have recently been studied as an attractive source for the development of a bioartificial liver (BAL). Here we evaluate the differentiation capacity of hES cells into hepatocytes. hES cells were subjected to suspension culture for 5 days, and then cultured onto poly-amino-urethane (PAU)-coated, nonwoven polytetrafluoroethylene (PTFE) fabric in the presence of fibroblast growth factor-2 (bFGF) (100 ng/ml) for 3 days, then with deleted variant of hepatocyte growth factor (dHGF) (100 ng/ml) and 1% dimethyl sulfoxide (DMSO) for 8 days, and finally with dexamethasone (10(-7) M) for 3 days. The hES cells showed gene expression of albumin in a time-dependent manner of the hepatic differentiation process. The resultant hES-derived hepatocytes metabolized the loaded ammonia and lidocaine at 7.8% and 23.6%, respectively. A million of such hepatocytes produced albumin and urea at 351.2 ng and urea at 7.0 microg. Scanning electron microscopy showed good attachment of the cells on the surface of the PTFE fabric and well-developed glycogen rosettes and Gap junction. In the present work we have demonstrated the efficient differentiation of hES cells to functional hepatocytes. The findings are useful to develop a BAL.
authors.affiliation: Department of Surgery, Okayama University Graduate School of Medicine and Dentistry, 2-5-1 Shikata-cho, Okayama 700-8558, Japan.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Soto-Gutierrez
authors.forename: Nalú
authors.initials: N
authors.lastname: Navarro-Alvarez
authors.forename: Jorge David
authors.initials: JD
authors.lastname: Rivas-Carrillo
authors.forename: Yong
authors.initials: Y
authors.lastname: Chen
authors.forename: Tomoki
authors.initials: T
authors.lastname: Yamatsuji
authors.forename: Noriaki
authors.initials: N
authors.lastname: Tanaka
authors.forename: Naoya
authors.initials: N
authors.lastname: Kobayashi
mesh_terms: D000199:Actins / Q000378:metabolism; D000418:Albumins / Q000235:genetics / Q000378:metabolism; D000641:Ammonia / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002455:Cell Division / Q000187:drug effects; D020099:Coated Materials, Biocompatible / Q000737:chemistry / Q000494:pharmacology; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism; D020869:Gene Expression Profiling; D017228:Hepatocyte Growth Factor / Q000235:genetics / Q000494:pharmacology*; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D008012:Lidocaine / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D011073:Polyamines / Q000737:chemistry; D011138:Polytetrafluoroethylene / Q000737:chemistry / Q000494:pharmacology; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D013784:Textiles; D013997:Time Factors; D014508:Urea / Q000378:metabolism; D014520:Urethane / Q000737:chemistry
chemical_list: D000199:Actins; D000418:Albumins; D020099:Coated Materials, Biocompatible; D011073:Polyamines; D014520:Urethane; D017228:Hepatocyte Growth Factor; D000641:Ammonia; D014508:Urea; D011138:Polytetrafluoroethylene; D008012:Lidocaine