title: Chemically fixed nurse cells for culturing murine or primate embryonic stem cells.
abstract: In current and past practice, murine or primate embryonic stem (ES) cells are usually cultured on live nurse cells for growth that keeps the cells in an undifferentiated state. It is troublesome, however, to prepare nurse cells for each cell culture and it is difficult to completely remove the nurse cells when they are transferred. In this study, mouse and monkey ES cells were therefore grown on chemically fixed mouse embryonic fibroblast (MEF) or human amniotic epithelial (HAE) cells. MEF cells were fixed by incubation in a glutaraldehyde or formaldehyde solution. HAE cells were immortalized by transfection of hTERT and chemically fixed with the same reagents. When mouse ES cells were cultured on these chemically fixed cells, the mouse ES cells grew well and expressed alkaline phosphatase, SSEA-1, and Oct-3/4 as their markers, indicating their undifferentiated state. The monkey ES cells also grew well and expressed alkaline phosphatase, SSEA-4, and Oct-4 as their markers, indicating their undifferentiated state. Freeze-drying HAE or MEF cells did not change their ability to support the undifferentiated growth of ES cells. Additionally, the chemically fixed cells could be utilized repeatedly in the culture of ES cells. These results demonstrate that chemically fixed nurse cells are useful for the maintenance of ES cells in an undifferentiated state in culture.
authors: Regenerative Medical Bioreactor Project, Kanagawa Academy of Science and Technology, KSP East 309, 3-2-1 Sakado, Takatsu-ku, Kawasaki, Kanagawa 213-0012, Japan. y-ito@riken.jp
authors: Yoshihiro
authors: Y
authors: Ito
authors: Makiko
authors: M
authors: Kawamorita
authors: Toshio
authors: T
authors: Yamabe
authors: Toru
authors: T
authors: Kiyono
authors: Kanji
authors: K
authors: Miyamoto
mesh_terms: D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D018920:Coculture Techniques; D053595:Embryonic Stem Cells / Q000737:chemistry / Q000166:cytology / Q000502:physiology*; D004847:Epithelial Cells / Q000737:chemistry* / Q000187:drug effects; D005347:Fibroblasts / Q000737:chemistry* / Q000187:drug effects; D005557:Formaldehyde / Q000494:pharmacology; D005976:Glutaral / Q000494:pharmacology; D000882:Haplorhini; D006801:Humans; D016256:Lewis X Antigen / Q000032:analysis; D051379:Mice; D016853:Muromonab-CD3 / Q000032:analysis; D019098:Telomerase / Q000235:genetics
chemical_list: D016256:Lewis X Antigen; D016853:Muromonab-CD3; D005557:Formaldehyde; D019098:Telomerase; D000469:Alkaline Phosphatase; D005976:Glutaral