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New Strategy for Manufacturing of Well-Orderd Tissue and Organ

制造有序组织和器官的新策略

基本信息

批准号：
15360436
负责人：
SHINKAI Masashige
金额：
$ 9.86万
依托单位：
THE UNIVERSITY OF TOKYO
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

项目摘要

I.Arrangement of exthacellular matrix using Transglutaminase gelathon method and design of basic frame peptideSome peptides of a basic frame of the matrix including the consensus sequence of the cell attachment factor such as the RGD peptide and active site to the Transglutaminase (TGase) was designed and prepared. These several kinds of peptide were mixed with TGase and spread on the cell arranged on the plate. The artificial matrix was arranged in intercellular. Properties of the matrix changed depending on the amount of the RGD consensus sequence, and the material that changed the response to the cell could be developed.II.Formation of true three-dimensional structure using Transglutaminase gelation methodWhen BAM (Biocompatible Anchor for Membrane) was spread on the plate in an appropriate pattern and cell suspension was added, the immobilization of the cells was confirmed. Moreover, the cell could be made attached on the sidewall in the micro channel made with PDMS polymer. On the basis of such pseudo three-dimensional stricture, formation of a three-dimensional structure using the TGase gelation method was investigated. The rat hepatic parenchymal cells were arranged on the BAM pattern, and their albumin secretion was confirmed.III.Formation of true three-dimensional structure including heterostructureThe formation of the luminal structure such as blood vessels and the bile ducts was investigated. The micro channel structure of the gelatin gel was prepared using the mold of the channel with PDMS polymer. The fibroblast cells were included into this. Inlet and outlet tubes were connected to the micro channel structure and the medium was supplied into this. As a result, the cell in the structure could be maintained at least for a week. In addition, a complex three-dimensional structure was constructed with the arrangement of the HeLa cell and vascular endothelial cells on the surface of the gelatin gel and folding of this.

1.谷氨酰胺转氨酶明胶凝胶法排列细胞外基质及碱性框架肽的设计：设计并制备了一些基质基本框架的多肽，包括RGD肽和转谷氨酰胺酶(TGase)活性部位等细胞黏附因子的共同序列。这几种多肽与TGase混合，铺展在平板上排列的细胞上。人工基质呈胞间排列。使用转谷氨酰胺酶凝胶法形成真正的三维结构当BAM(生物相容性膜锚)以适当的图案铺展在平板上并加入细胞悬浮液时，确认了细胞的固定化。此外，在由PDMS聚合物制成的微通道中，可以将细胞附着在侧壁上。在这种伪三维狭窄的基础上，研究了利用TGase凝胶法形成三维结构的方法。大鼠肝实质细胞排列在BAM模式上，并确认其白蛋白分泌。III.包括异质结构在内的真三维结构的形成观察血管和胆管等管腔结构的形成。以聚二甲基硅氧烷(PDMS)为模板，制备明胶凝胶微通道结构。成纤维细胞被纳入其中。入口和出口管连接到微通道结构，介质被供应到该微通道结构中。因此，结构中的细胞可以保持至少一周。此外，还构建了HeLa细胞和血管内皮细胞在明胶表面的排列和折叠的复杂三维结构。