1988 Fiscal Year Final Research Report Summary
Development of matrix-culture system for osteoblastic cells using active-form ascorbic acid
| Project/Area Number |
62870073
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Functional basic dentistry
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| Research Institution | Hokkaido University |
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Principal Investigator |
KUBOKI Yoshinori Hokkaido University, School of Dentistry, 歯学部, 教授 (00014001)
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| Co-Investigator(Kenkyū-buntansha) |
TAKITA Hiroko Hokkaido University, School of Dentistry, 歯学部, 教務職員 (30125330)
FUJISAWA Ryuichi Hokkaido University, School of Dentistry, 歯学部, 助手 (40190029)
MIZUNO Morimichi Hokkaido University, School of Dentistry, 歯学部, 助手 (10125354)
HATA Ryuichiro Tokyo Medical and Dental University, Institute Medical Research, 難治疾患研究所, 講師 (10014276)
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| Project Period (FY) |
1987 – 1988
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| Keywords | Osteoblast / Osteogenic cells / Collagen gel / Matrix culture / 活性持続型アスコルビン酸 |
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| Research Abstract |
Osteogenesis is a complex process containing several kinds of cells, which interact each other with growth factors or differentiation factors. In vitro cell culture system is a useful tool for the research on this process. We attempted to establish a large-scale culture system of osteoblast-like cells. A clonal oste*oblast-like cell line (of mouse), MC3T3-E1, was used as a material. As a preliminary study, the cell line was cultured on convensional plastic dishes. The cells proliferated and were stacked in mult-layers, then, formed noduli and finally calcified. The active-form ascorbate was usefui in sustaining cell growth under serum dificient condition.
The cells were cultured on spherical beads of collagen gel having a diameter of 0.5-1.0 mm. The cells proliferated and covered the entire surface of the beads forming at most 7 cellular layers.
The cells were cultured on collagen gels cross-linked with 4 cross-linking agents. Among these agents, hexamethylenediisocyanate ( HMDIC ) and a long chain epoxy agent were found to be effective to prepare uncontracted gels for culture. The cells pro-liferated and were calcified on the gels.
The third method was to culture cells embeded in collagen gels. Under this condition, the cells proliferated and exhibited to increase alkaline phosphatase activity, which is a characteristics of osteoblastic cells. When guanidin/ HCL extracts of bone, crude bone morphogenetic proteins, were incorporated in this system, the cells responded with significant increase in alkaline phosphatase activity.
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Research Products
(14 results)
