Differentiation of human embryonic stem cells to hepatocytes using deleted variant of HGF and poly-amino-urethane-coated nonwoven polytetrafluoroethylene fabric.
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.
Soto-Gutierrez
Alejandro
A
Department of Surgery, Okayama University Graduate School of Medicine and Dentistry, 2-5-1 Shikata-cho, Okayama 700-8558, Japan.
Navarro-Alvarez
Nalú
N
Rivas-Carrillo
Jorge David
JD
Chen
Yong
Y
Yamatsuji
Tomoki
T
Tanaka
Noriaki
N
Kobayashi
Naoya
N
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
D000199:Actins; D000418:Albumins; D020099:Coated Materials, Biocompatible; D011073:Polyamines; D014520:Urethane; D017228:Hepatocyte Growth Factor; D000641:Ammonia; D014508:Urea; D011138:Polytetrafluoroethylene; D008012:Lidocaine