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[利用胶原玻璃膜优势开发新型细胞培养系统]

[Development of novel cell culture systems utilizing the advantages of collagen vitrigel membrane].

作者信息

Takezawa Toshiaki, Fukuda Maya, McIntosh-Ambrose Winnette, Ko Ji-Ae, Elisseeff Jennifer, Haga Sanae, Ozaki Michitaka, Kato Kiyoko, Wang Pi-Chao, Uchino Tadashi, Nishida Teruo

机构信息

Transgenic Animal Research Center, National Institute of Agrobiological Sciences, Ibaraki, Japan.

出版信息

Yakugaku Zasshi. 2010 Apr;130(4):565-74. doi: 10.1248/yakushi.130.565.

Abstract

The white of an egg, rendered opaque by boiling, can be converted into a thin, transparent and rigid material like glass by evaporating the moisture. This phenomenon is known as the vitrification of heat-denatured proteins. We applied vitrification technology to a collagen gel and converted it into a rigid glass-like material. We attempted to rehydrate the glass-like material and succeeded in preparing a novel stable state of collagen gel that was a thin and transparent membrane with excellent gel strength and protein permeability. We called it "collagen vitrigel" because it was produced from the vitrification process of a traditional hydrogel. Further, a framework-embedded collagen vitrigel membrane that can be easily turned inside out with tweezers was prepared by inserting a nylon membrane ring in the collagen sol prior to the gelation, thereby allowing the membrane to function as a removable cell culture substratum. Different types of anchorage-dependent cells could be cultured on both surfaces of the substratum by the manipulation of two-dimensional cultures, and consequently a three-dimensional crosstalk model with paracrine effects from each cell type was reconstructed. Also, the collagen vitrigel membrane containing a bioactive molecule provided a drug delivery system (DDS) with sustainable release. In this review, we summarize the recent progress of applied studies using the collagen vitrigel membrane as follows: a corneal model for eye irritant and permeability tests, a skin model for sensitization test, a renal glomerular model for evaluating blood filtration, an endometrial model for developing a new treatment and a DDS of hepatocyte growth factor for improving liver disorder.

摘要

煮过之后变得不透明的蛋清,通过水分蒸发可转化为一种像玻璃一样薄、透明且坚硬的材料。这种现象被称为热变性蛋白质的玻璃化。我们将玻璃化技术应用于胶原蛋白凝胶,并将其转化为一种坚硬的玻璃状材料。我们试图使这种玻璃状材料再水化,并成功制备出一种新型的稳定状态的胶原蛋白凝胶,它是一种具有优异凝胶强度和蛋白质渗透性的薄而透明的膜。我们将其称为“胶原蛋白玻璃凝胶”,因为它是由传统水凝胶的玻璃化过程产生的。此外,通过在凝胶化之前将尼龙膜环插入胶原蛋白溶胶中,制备了一种可以用镊子轻松翻转的框架嵌入型胶原蛋白玻璃凝胶膜,从而使该膜能够作为一种可移除的细胞培养基质发挥作用。通过二维培养的操作,不同类型的贴壁依赖性细胞可以在该基质的两个表面上进行培养,因此重建了一个具有每种细胞类型旁分泌效应的三维串扰模型。而且,含有生物活性分子的胶原蛋白玻璃凝胶膜提供了一种具有可持续释放功能的药物递送系统(DDS)。在这篇综述中,我们总结了使用胶原蛋白玻璃凝胶膜的应用研究的最新进展如下:用于眼刺激和渗透性测试的角膜模型、用于致敏测试的皮肤模型、用于评估血液过滤的肾小球模型、用于开发新疗法的子宫内膜模型以及用于改善肝脏疾病的肝细胞生长因子的药物递送系统。

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