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Name (Title):
Guoping Chen (Group Leader) and Naoki Kawazoe (Researcher)
Affiliation:
International Center for Materials Nanoarchitectonics and Biomaterials
Center, National Institute for Materials Science
Address:
1-1 Namiki, Tsukuba, Ibaraki 3050044, Japan
Email: Guoping.CHEN@nims.go.jp
Home Page: http://www.nims.go.jp/bmc/
Presentation Title:
Manipulation of Stem Cell Functions by Patterned Polymer Surfaces
Abstract:
Manipulation of stem cell functions is an important technique for tissue engineering. It was
realized by patterned surfaces. Gelatin was pattern-grafted on cell-culture polystyrene plate
surface and used for culture of human mesenchymal stem cells (MSCs). The pattern was prepared
by UV-irradiating the photoreactive azidobenzoyl-derivatized gelatin-coated surface of a
polystyrene plate through a photomask with 200-µm-wide stripes. The MSCs adhered on both
polystyrene and gelatin-grafted surfaces, proliferated to reach confluent. The cells were stained
with alkaline phosphatase (ALP) and alizarin red S (calcium staining) to analyze the osteogenic
differentiation of MSCs on these surfaces. The cells on both polystyrene and gelatin-grafted
surfaces were positively stained by alkaline phophatase. No pattern stain of alkaline phosphatase
was detected. However, the cells on gelatin-grafted surface were strongly stained by alizarin red
S after two weeks culture, while cells on polystyrene surface very weakly stained after two weeks
culture. Therefore the alizarin red S staining followed the gelatin pattern. The pattern became less
distinct after three weeks culture, and not evident after four weeks culture. The MSCs showed
osteogenic differentiation pattern following the gelatin pattern in the first three weeks. Gene
expression analyses using real-time PCR indicated that MSCs cultured on the gelatin-grafted
surface showed higher level of genes encoding alkaline phosphatase and bone sialoprotein than
did on polystyrene surface. The expression of gene encoding osteocalcin was at almost the same
level for both surfaces. These results suggest that the osteogenic differentiation rate of MSCs on
the polystyrene and gelatin-grafted surfaces were different, and that the MSCs differentiated
more quickly on the gelatin-grafted surface than did on the polystyrene surface. The pattern
surface could be used to control differentiation of stem cells, and may be used to reconstruct the
complex structure of tissue and organs.
12
a b c d
Oral Presentation 12
Fig. 1. Photomicrographs of photomask (a), PVA-patterned polystyrene surface (b), and MSCs
cultured on the PVA-patterned surface in osteogenic induction medium for 2 weeks without staining
(c) and stained with alizarin red S (d).
References:
1. G. Chen, N. Kawazoe, Y. Fan, Y. Ito, T. Tateishi, Langmuir, 23 (2007) 5864.
2. L. Guo, N. Kawazoe, T. Hoshiba, T. Tateishi, G. Chen, X. Zhang, Journal of Biomedical
Materials Research: Part A, 87(2008) 903.
3. N. Kawazoe, L. Guo, M. J. Wozniak, Y. Imaizumi, T. Tateishi , X. Zhang, G. Chen, Journal of
Nanoscience and Nanotechnology, 9 (2009) 230.