1994 Fiscal Year Final Research Report Summary
Development of image analyze systems of gene expression in co-cultured cells
| Project/Area Number |
05557042
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Circulatory organs internal medicine
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| Research Institution | Osaka University |
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Principal Investigator |
KUZUYA Tsunehiko Osaka Univ., Medical School., Associate Prof., 医学部, 助教授 (80150340)
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| Co-Investigator(Kenkyū-buntansha) |
NISHIDA Masashi Osaka Univ., Medical School., Resident, 医学部・附属病院, 医員
HOSHIDA Shiro Osaka Univ., Medical School., Associate Prof., 医学部, 助手 (80238732)
HORI Masatsugu Osaka Univ., Medical School., Associate Prof., 医学部, 講師 (20124779)
TADA Michihiko Osaka Univ., Medical School., Professor, 医学部, 教授 (90093434)
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| Project Period (FY) |
1993 – 1994
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| Keywords | Cardiac myocyte / gene expression / image analysis / calcium |
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| Research Abstract |
The purpose of this study was to establish co-culture system of heart cells under microscopy and analyze gene expression and function of the cells keeping inter-cellular interaction.
1. Cell culture
We isolated and cultured cardiac myocytes and coronary microvascular endothelial cells from neonatal and adult rat hearts. These cells were (co-) cultured on glass chamber slide and their phenotype was observed under microscopy as in 3.
2. Gene transfer
We tried to transfer gene constructs into primary cultures of heart cells. Instead of direct injection which was not budgeted, we modified calcium phosphate method and lipofection method to obtain good transfer efficiency.
3. Optic system
Inverted fluorescence microscopy system with fluorescence and luminescence detectors was attached to a stage with superfusion system. Cultured cells were placed on the stage and their phenotype (fluorescence) and genotype (chemiluminescence) was examined.
Due to the difference of optical characteristics between fluorescence and luminescence, simultaneous measurement of both property was not achieved. To analyze time course of these measures, a memory board is now installed in the system.
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