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Basic research for cell-material interaction by using molecularly controlled surface and gene expression analysis

利用分子控制表面和基因表达分析进行细胞-材料相互作用的基础研究

基本信息

批准号：
09480255
负责人：
SERIZAWA Takeshi
金额：
$ 6.27万
依托单位：
Kagoshima University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

项目摘要

The adhesion and function of HeLa S3 cells on lipid films were investigated. The function of the cells adhered to lipid films was estimated by HSP70B and HSP47 mRNA expression using a reverse transcription-polymerase chain reaction (RT-PCR) analysis. It is apparent that HeLa S3 cells could adhere to lipid films as well as tissue culture poly(styrene)(TCPS) HSPs mRNA expression on the lipid film depended on each lipid films. HeLa S3 cells adhered to the lipid films of DPPC, DPPE and DPPS expressed the same level HSPs mRNA as TCPS. On the other hand, high HSPs mRNA expressions were observed in the HeLa S3 cells fastened to the lipid films of synthetic lipids. The serum proteins effected the HSPs expression in the case of the synthetic lipids. These results imply that cell-material interactions were not simple, and that the mRNA study was a powerful tool in the investigation of cell functions, especially concerning the reaction of weak stimulus such as cell-material contact.In order to interpret how cells recognize biomaterials, nucleic factor-kappa B(NF-ィイD2κィエD2B) activation in the attached HeLa S3 cells on various substrates was evaluated. As substrates, materials of hydrophilic nature (cellulose, poly(acrylamide) grafted poly(ethylene)(PAAm-g-PE), and lipids films) were used. The contemporary assay method for NF-ィイD2κィエD2B was modified to fit our system. As a result, NF-ィイD2κィエD2B activation varied depending on the substrates. The NF-ィイD2κィエD2B outcome was induced significantly in the HeLa S3 cells that had adhered onto the lipid films in a short time. On the other hand, high levels of NF-ィイD2κィエD2B induction was observed in the HeLa cells adhered to the celluose and PAAm-g-PE after a 24 hour incubation period. The induction of NF-ィイD2κィエD2B by cell-material seemed to be closely related to the biocompatibility of the material.

研究了 HeLa S3 细胞在脂质膜上的粘附和功能。使用逆转录聚合酶链式反应 (RT-PCR) 分析，通过 HSP70B 和 HSP47 mRNA 表达来评估粘附在脂质膜上的细胞的功能。很明显，HeLa S3 细胞可以粘附在脂膜上，并且组织培养物聚（苯乙烯）（TCPS）HSP mRNA 在脂膜上的表达取决于每个脂膜。粘附在DPPC、DPPE和DPPS脂质膜上的HeLa S3细胞表达与TCPS相同水平的HSPs mRNA。另一方面，在固定于合成脂质的脂质膜上的 HeLa S3 细胞中观察到高 HSP mRNA 表达。在合成脂质的情况下，血清蛋白影响热休克蛋白的表达。这些结果意味着细胞-材料相互作用并不简单，mRNA研究是研究细胞功能的有力工具，特别是涉及细胞-材料接触等弱刺激的反应。为了解释细胞如何识别生物材料，评估了附着在各种基质上的HeLa S3细胞中的核酸因子-κB（NF-ィイD2κエD2B）激活。使用亲水性材料（纤维素、聚丙烯酰胺接枝聚乙烯（PAAm-g-PE）和脂质膜）作为基材。 NF-ィイD2κィエD2B 的当代测定方法经过修改以适合我们的系统。结果，NF-ィイD2κィエD2B 的激活根据底物的不同而变化。在短时间内粘附到脂质膜上的 HeLa S3 细胞中显着诱导了 NF-ィイD2κィエD2B 结果。另一方面，在 24 小时孵育期后，在粘附于纤维素和 PAAm-g-PE 的 HeLa 细胞中观察到高水平的 NF-ィイD2κィD2B 诱导。细胞材料诱导NF-ィイD2κエD2B似乎与材料的生物相容性密切相关。