Epithelial-mesenchymal transition in rhesus monkey embryonic stem cell colonies: the role of culturing conditions

Colonies of rhesus monkey embryonic stem cells (rhESC; cell line R366.4) have been described before to show a spatially ordered process of epithelial-mesenchymal transition in vitro. In the present investigations, we have studied variables of culturing conditions which influence the reproducibility...

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Bibliographic Details
Published in:In vitro cellular & developmental biology. Animal 2010-06, Vol.46 (6), p.516-528
Main Authors: Maranca-Hüwel, Birgit, Denker, Hans-Werner
Format: Article
Language:English
Subjects:
Actins
Age
Animal Genetics and Genomics
Animals
Biomedical and Life Sciences
CELL AND TISSUE MODELS
Cell Biology
Cell Culture
Cell Culture Techniques
Cell Differentiation
Cell lines
Cells, Cultured
Colonies
Cultured cells
Developmental Biology
Embryo cells
Embryo fibroblasts
Embryo, Mammalian - metabolism
Embryonic stem cells
Embryonic Stem Cells - cytology
Embryonic Stem Cells - metabolism
Embryos
Enzymes
Epithelial Cells - cytology
Epithelial Cells - metabolism
Feeder cells
Fibroblasts - cytology
Fibroblasts - metabolism
Gastrulation
Irradiation
Life Sciences
Macaca mulatta
Mesenchymal Stromal Cells - cytology
Mesenchymal Stromal Cells - metabolism
Mice
Mitomycin C
Primates
Radiation
Stem Cells
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Summary:Colonies of rhesus monkey embryonic stem cells (rhESC; cell line R366.4) have been described before to show a spatially ordered process of epithelial-mesenchymal transition in vitro. In the present investigations, we have studied variables of culturing conditions which influence the reproducibility of the formation of crater-like ingression centers in the colonies. Critical parameters are found to be age and density of mouse embryonic fibroblast (MEF) feeder cell layers, the mode of mitotic inactivation of the MEFs (mitomycin C, or irradiation), and the mode of rhESC isolation during subculturing (enzymatic/mechanical cell cluster isolation; type of enzyme). The described culturing system appears to offer a reproducible in vitro model potentially useful for studies on cellular processes involved in gastrulation in the primate.
ISSN:1071-2690
1543-706X
DOI:10.1007/s11626-010-9272-8