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Development of novel intelligent tissue culture surfaces to accelerate cell proliferation and recover cell sheet under serum free conditions, using physiologically active peptides

使用生理活性肽开发新型智能组织培养表面，以加速细胞增殖并在无血清条件下恢复细胞片层

基本信息

批准号：
16200036
负责人：
OKANO Teruo
金额：
$ 32.12万
依托单位：
Tokyo Woman's Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2006
项目状态：
已结题

项目摘要

In order to evolve advanced method of cell sheet engineering, we have developed novel temperature responsive polymers grafted tissue culture polystyrene (TCPS), copolymerizing 2-carboxyisopropylacrylamide monomer as following immobilization of physiologically active peptides with IPAAm monomer (PIPAAm-CIPAAm-TCPS). The novel culture surfaces, PIPAAm-CIPSSm-TCPS, modified with RGD peptides attain culturing cells even under serum free conditions, and cells were successfully recovered as a sheet from the surfaces by lowering temperature. Cell culture onto the novel surfaces is useful for cells to avoid contamination of prion protein and virus during cell culture due to serum free condition. RGD and PHSRN modified surface exhibited more cell adhesive property than only RDG. In contrast, introduction of RDG and Insulin onto the novel culture surface achieve acceleration of proliferation of cultured cells. RGD peptides I inked with biotin were introduced onto avidin proteins modified onto the novel surfaces. Resulting RGD modified surfaces also showed cell adhesion and proliferation properties even under serum free conditions, strongly suggesting various physiologically active peptides, which contain biotin component, are easily immobilized onto the novel surfaces modified with avidin. Determination of appropriate physiologically active peptides for corneal epithelium cells, retinal pigment epithelium cells, alveolar cells and cardiac muscle cells may promote clinical application utilizing these kinds of cell sheets.

为了发展先进的细胞片工程方法，我们开发了新型温度响应性聚合物接枝组织培养聚苯乙烯（TCPS），将2-羧基异丙基丙烯酰胺单体共聚为IPAAm单体固定生理活性肽（PIPAAm-CIPAAm-TCPS）。用RGD肽修饰的新型培养表面PIPAAm-CIPSSm-TCPS即使在无血清条件下也能培养细胞，并且通过降低温度从表面成功地回收细胞作为薄片。在新型表面上的细胞培养对于细胞是有用的，以避免由于无血清条件而在细胞培养期间污染朊病毒蛋白和病毒。RGD和PHSRN修饰的表面比单独的RDG具有更好的细胞粘附性。相比之下，将RDG和胰岛素引入到新型培养表面上实现了培养细胞增殖的加速。将生物素连接的RGD肽I引入到修饰在新型表面上的亲和素蛋白上。所得的RGD修饰的表面也显示出细胞粘附和增殖性能，即使在无血清条件下，强烈建议各种生理活性肽，其中包含生物素组分，很容易固定到新的表面上的亲和素修饰。确定角膜上皮细胞、视网膜色素上皮细胞、肺泡细胞和心肌细胞的适当生理活性肽可促进利用这些种类的细胞片的临床应用。