title: Mouse embryonic stem cells cultured under serum- and feeder-free conditions maintain their self-renewal capacity on hydroxyapatite.
abstract: New tissue engineering techniques based on embryonic stem (ES) cells and artificial scaffolds are required for regenerative medicine. Because artificial scaffolds can regulate the differentiation states of ES cells, compatibility between the cells and artificial scaffolds is important. To our knowledge, this study is the first report showing that mouse ES (mES) cells can be maintained in undifferentiated state on hydroxyapatite coated with gelatin. In contrast to previous studies, our culture medium was serum-free and included a GSK-3 inhibitor. Under these conditions, mES colony morphology was similar to that of an undifferentiated state; mES cells expressed the pluripotent-specific factors Oct-3/4 and Nanog, and they maintained the ability to differentiate into the three germ layers. Moreover, a GSK-3 inhibitor blocked the expression of integrin subunits that bind to laminin which are known to induce the differentiation of mES cells. These findings indicate that mES cells can be cultured under serum- and feeder-free conditions and maintained in an undifferentiated state on a composite with hydroxyapatite and that this composite can be used to control the differentiation of stem cells.
authors.affiliation: Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10 Kanda-Surugadai, Chiyoda, Tokyo 101-0062, Japan. Electronic address: nag-bcr@tmd.ac.jp.
authors.forename: Akiko
authors.initials: A
authors.lastname: Nagai
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Hattori
authors.forename: Michiko
authors.initials: M
authors.lastname: Hirose
authors.forename: Atsuo
authors.initials: A
authors.lastname: Ogura
authors.forename: Kosuke
authors.initials: K
authors.lastname: Nozaki
authors.forename: Mamoru
authors.initials: M
authors.lastname: Aizawa
authors.forename: Kimihiro
authors.initials: K
authors.lastname: Yamashita
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media / Q000494:pharmacology; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D017886:Durapatite / Q000494:pharmacology*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure; D061252:Feeder Cells / Q000166:cytology* / Q000187:drug effects; D018398:Homeodomain Proteins / Q000378:metabolism; D007211:Indoles / Q000494:pharmacology; D016023:Integrins / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D000071317:Nanog Homeobox Protein; D010091:Oximes / Q000494:pharmacology; D017550:Spectroscopy, Fourier Transform Infrared; D014961:X-Ray Diffraction
chemical_list: C483321:6-bromoindirubin-3'-oxime; D015415:Biomarkers; D003470:Culture Media; D016895:Culture Media, Serum-Free; D018398:Homeodomain Proteins; D007211:Indoles; D016023:Integrins; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D010091:Oximes; D017886:Durapatite; D000469:Alkaline Phosphatase
keywords: Composite; GSK-3 inhibitor; Hydroxyapatite; Integrin; Mouse embryonic stem cell; Scaffold