Hierarchical Micro-Nano Surface Topography Promotes Long-Term Maintenance of Undifferentiated Mouse Embryonic Stem Cells.
Understanding of stem cell-surface interactions and, in particular, long-term maintenance of stem cell pluripotency on well-defined synthetic surfaces is crucial for fundamental research and biomedical applications of stem cells. Here, we show that synthetic surfaces possessing hierarchical micro-nano roughness (MN-surfaces) promote long-term self-renewal (>3 weeks) of mouse embryonic stem cells (mESCs) as monitored by the expression levels of the pluripotency markers octamer-binding transcription factor 4 (Oct4), Nanog, and alkaline phosphatase. On the contrary, culturing of mESCs on either smooth (S-) or nanorough polymer surfaces (N-surfaces) leads to their fast differentiation. Moreover, we show that regular passaging of mESCs on the hierarchical MN-polymer surface leads to an increased homogeneity and percentage of Oct4-positive stem cell colonies as compared to mESCs grown on fibroblast feeder cells. Immunostaining revealed the absence of focal adhesion markers on all polymer substrates studied. However, only the MN-surfaces elicited the formation of actin-positive cell protrusions, indicating an alternative anchorage mechanism involved in the maintenance of mESC stemness.
Karlsruhe Institute of Technology (KIT) , Department of Cell- and Neurobiology, Zoological Institute, Haid-und-Neu-Straße 9, 76131 Karlsruhe, Germany.|Karlsruhe Institute of Technology (KIT) , Institute of Functional Interfaces (IFG), PO Box 3640, 76021 Karlsruhe, Germany.
Mona
M
Jaggy
Karlsruhe Institute of Technology (KIT) , Institute of Toxicology and Genetics (ITG), PO Box 3640, 76021 Karlsruhe, Germany.
Ping
P
Zhang
Karlsruhe Institute of Technology (KIT) , Department of Cell- and Neurobiology, Zoological Institute, Haid-und-Neu-Straße 9, 76131 Karlsruhe, Germany.
Alexandra M
AM
Greiner
Karlsruhe Institute of Technology (KIT) , Department of Cell- and Neurobiology, Zoological Institute, Haid-und-Neu-Straße 9, 76131 Karlsruhe, Germany.|Karlsruhe Institute of Technology (KIT) , Institute of Functional Interfaces (IFG), PO Box 3640, 76021 Karlsruhe, Germany.
Tatjana J
TJ
Autenrieth
Karlsruhe Institute of Technology (KIT) , Institute of Toxicology and Genetics (ITG), PO Box 3640, 76021 Karlsruhe, Germany.
Victoria
V
Nedashkivska
Karlsruhe Institute of Technology (KIT) , Institute of Toxicology and Genetics (ITG), PO Box 3640, 76021 Karlsruhe, Germany.
Alexander N
AN
Efremov
Karlsruhe Institute of Technology (KIT) , Institute of Toxicology and Genetics (ITG), PO Box 3640, 76021 Karlsruhe, Germany.
Christine
C
Blattner
Karlsruhe Institute of Technology (KIT) , Department of Cell- and Neurobiology, Zoological Institute, Haid-und-Neu-Straße 9, 76131 Karlsruhe, Germany.|Karlsruhe Institute of Technology (KIT) , Institute of Functional Interfaces (IFG), PO Box 3640, 76021 Karlsruhe, Germany.
Martin
M
Bastmeyer
Karlsruhe Institute of Technology (KIT) , Institute of Toxicology and Genetics (ITG), PO Box 3640, 76021 Karlsruhe, Germany.|Karlsruhe Institute of Technology (KIT) , Institute of Organic Chemistry, PO Box 3640, 76021 Karlsruhe, Germany.
Pavel A
PA
Levkin
D000818:Animals; D002454:Cell Differentiation; D053595:Embryonic Stem Cells / Q000166:cytology*; D051379:Mice; D008855:Microscopy, Electron, Scanning; D013499:Surface Properties
mouse embryonic stem cells; polymer surface; stemness maintenance; surface roughness; synthetic stem cell substrate