Development of Cultured Renal Tubules with Cell Sheet Engineering

利用细胞片工程开发培养肾小管

• 批准号: 13680954
• 负责人: YAMATO Masayuki
• 金额: $ 2.56万
• 依托单位: Tokyo Women's Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13680954/
• 关键词: poly(N-isopropylacrylamide); temperature-responsive surface; cell sheet manipulation; kidney epithelial cell; urinary tubule; bioartificial kidney

We developed a temperature-responsive cultured dishes by covalent grafting of poly (N-isopropylacrylamide) which shows the hydrophilic/hydrophobic change of surface property. This surface shows weak hydrophobicity similarly to commercial tissue culture polystyrene dishes and allows various cell adhesion and spreading on it at 37℃. But the cells are detached from the highly hydrated surfaces without any need of proteolytic enzymes such as trypsin after reducing temperature below 32℃. All the spread cells connecting via cell-cell junctions are harvested as a single contiguous cell sheet. In the present study, we examined the utilization of harvested kidney epithelial cell sheets in the development of bioartificail renal tubules by combining the cell sheets and porous membranes for dialysis in terms of molecular transport and production. We developed a non-invasive cell sheet manipulation method from culture dishes to other surfaces such as porous membranes. Experimental devices were prepared by combining of primary human kidney epithelial cells and porous membranes. The expression and localization of various cell surface proteins are confirmed by immunostaining and confocal laser scanning microscopy.

我们通过共价（N-异丙基丙烯酰胺）的共价嫁接来开发温度响应性培养的培养皿，该菜肴显示了表面特性的亲水/疏水性变化。该表面显示出类似于商业组织培养的聚苯乙烯菜肴的弱疏水性，并允许各种细胞粘附并在37℃上扩散。但是细胞从高度水合的表面分离，而无需蛋白水解酶，例如降低温度以下的胰蛋白酶。通过细胞 - 细胞连接连接的所有扩散细胞都作为单个连续的细胞片收获。在本研究中，我们通过将细胞表和多孔膜结合在分子运输和生产方面，研究了收获的肾上皮细胞表在生物肉质肾块茎开发中的利用。我们开发了一种非侵入性细胞片操纵方法，从培养皿到其他表面，例如多孔膜。通过结合原代人肾上皮细胞和多孔膜来制备实验装置。通过免疫染色和共聚焦激光扫描显微镜证实了各种细胞表面蛋白的表达和定位。

项目成果

Utsumi K: "Alteration of collagen IV in acutely deteriorated renal allografts"Transplantation. 71・12. 1757-1765 (2001)

Utsumi K：“急性恶化的同种异体移植肾中胶原蛋白 IV 的改变”71・12 1757-1765（2001）。

T.Shimizu, M.Yamato, T.Akatsu, T.SHibata, Y.Isoi, A.Kikuchi M.Umezu, T.Okano: "Electrically communicating three-dimensional cardiac tissue mimic fabricated by layered cultured cardiomyocyte sheets"J. Biomed. Mater. Res.. 60. 110-117 (2002)

T.Shimizu、M.Yamato、T.Akatsu、T.SHIbata、Y.Isoi、A.Kikuchi M.Umezu、T.Okano：“由分层培养的心肌细胞片制造的电通信三维心脏组织模拟物”J。

A. Kushida, M. Yamato, Y. Isoi, A. Kikuchi and T. Okano, Ed. Y. Ikada, Y. Umakoshi and T. Hotta: "Two-dimensional renal cell-sheet manipulation for a novel bioartificial renal tubule., Tissue Engineering for Therapeutic Use 6"123-128 (2002)

A. Kushida、M. Yamato、Y. Isoi、A. Kikuchi 和 T. Okano，Ed。

M.Yamato, C.Konno, M.Utsumi, A.Kikuchi, T.Okano: "Thermally responsive polymer-grafted surfaces facilitate patterned cell seeding and co-culture"Biomaterials. 23. 561-567 (2002)

M.Yamato、C.Konno、M.Utsumi、A.Kikuchi、T.Okano：“热响应聚合物接枝表面有利于图案化细胞接种和共培养”生物材料。

T.Shimizu, M.Yamato, Y.Isoi, T.Akutsu, T.Setomaru, K.Abe, A.Kikuchi, M.Umezu, T.: "Febrication of pulsatile cardiac tissue grafts using a novel 3-dimensional cell sheet manipulation technique and temperature-responsive cell culture surfaces"Circ. Res.. 90

T.Shimizu、M.Yamato、Y.Isoi、T.Akutsu、T.Setomaru、K.Abe、A.Kikuchi、M.Umezu、T.：“使用新型 3 维细胞片对脉动心脏组织移植物进行加热