Coating extracellular matrix proteins on a (3-aminopropyl)triethoxysilane-treated glass substrate for improved cell culture.
We demonstrate that a (3-aminopropyl)triethoxysilane-treated glass surface is superior to an untreated glass surface for coating with extracellular matrix (ECM) proteins when used as a cell culture substrate to observe cell physiology and behavior. We found that MDCK cells cultured on untreated glass coated with ECM removed the coated ECM protein and secreted different ECM proteins. In contrast, the cells did not remove the coated ECM protein when seeded on (3-aminopropyl)triethoxysilane-treated (i.e., silanized) glass coated with ECM. Furthermore, the morphology and motility of cells grown on silanized glass differed from those grown on non-treated glass, even when both types of glass were initially coated with laminin. We also found that cells on silanized glass coated with laminin had higher motility than those on silanized glass coated with fibronectin. Based on our results, we suggest that silanized glass is a more suitable cell culture substrate than conventional non-treated glass when coated by ECM for observations of ECM effects on cell physiology.
Division of Bioengineering and Bioinformatics, Graduate School of Information Science and Technology, Hokkaido University, Sapporo, Japan; Transdisciplinary Life Science Course, Faculty of Advanced Life Science, Hokkaido University, Sapporo, Japan.
Hiro-taka
HT
Masuda
Transdisciplinary Life Science Course, Faculty of Advanced Life Science, Hokkaido University, Sapporo, Japan.
Seiichiro
S
Ishihara
Department of Biomolecular Engineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan.
Ichiro
I
Harada
Transdisciplinary Life Science Course, Faculty of Advanced Life Science, Hokkaido University, Sapporo, Japan.
Takeomi
T
Mizutani
Department of Medical Physics, Graduate School of Medicine, Hokkaido University, Sapporo, Japan.
Masayori
M
Ishikawa
Transdisciplinary Life Science Course, Faculty of Advanced Life Science, Hokkaido University, Sapporo, Japan.
Kazushige
K
Kawabata
Transdisciplinary Life Science Course, Faculty of Advanced Life Science, Hokkaido University, Sapporo, Japan.
Hisashi
H
Haga
D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D002465:Cell Movement / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D020099:Coated Materials, Biocompatible / Q000737:chemistry* / Q000494:pharmacology; D003470:Culture Media / Q000737:chemistry; D004285:Dogs; D016326:Extracellular Matrix Proteins / Q000737:chemistry* / Q000494:pharmacology; D005898:Glass / Q000737:chemistry*; D061985:Madin Darby Canine Kidney Cells; D011437:Propylamines; D011554:Pseudopodia / Q000187:drug effects; D012821:Silanes / Q000737:chemistry*
D020099:Coated Materials, Biocompatible; D003470:Culture Media; D016326:Extracellular Matrix Proteins; D011437:Propylamines; D012821:Silanes; C477625:amino-propyl-triethoxysilane
(3-aminopropyl)triethoxysilane; MDCK cells; cell migration; culture substrate; extracellular matrix; fibronectin; glass; laminin