Haraguchi Kazutoshi, Takehisa Toru, Ebato Makiko
Material Chemistry Laboratory, Kawamura Institute of Chemical Research, 631 Sakado, Sakura, Chiba, 285-0078, Japan.
Biomacromolecules. 2006 Nov;7(11):3267-75. doi: 10.1021/bm060549b.
Cell cultivation on the surface of a novel poly(N-isopropylacrylamide) (PNIPA) hydrogel (N-NC gel), consisting of a specific type of organic (PNIPA)/inorganic (clay) network, was studied using three cell types; HepG2 human hepatoma cells, human dermal fibroblasts, and human umbilical vein endothelial cells. For the first time, it was found that cells could be cultured to be confluent on the surfaces of PNIPA hydrogels using N-NC gels, regardless of gel thickness. Cell adhesion and proliferation on N-NC gels exhibit strong dependencies on clay concentration (C(clay)), and the numbers of cultured cells are maximum at about C(clay) = 6 x 10(-2) mol (45.72 g)/1 L of H(2)O. On the contrary, it was almost impossible to culture cells on conventional, chemically crosslinked PNIPA hydrogels, regardless of their cross-linker concentration. The reasons why cells cultured only on the surfaces of N-NC gels with their specific network structure and composition were discussed in terms of water content, protein adsorption, surface flatness, hydrophobicity of dehydrated PNIPA chains, and the anionic charge on exfoliated clay. Finally, it was found that cells cultured on the surfaces of N-NC gels could be detached in the forms of sheets of cells without trypsin treatment, but by just decreasing the temperature to 20 degrees C.
使用三种细胞类型,即HepG2人肝癌细胞、人皮肤成纤维细胞和人脐静脉内皮细胞,研究了在一种新型聚(N-异丙基丙烯酰胺)(PNIPA)水凝胶(N-NC凝胶)表面的细胞培养情况,该水凝胶由特定类型的有机(PNIPA)/无机(粘土)网络组成。首次发现,使用N-NC凝胶时,无论凝胶厚度如何,细胞都可以在PNIPA水凝胶表面培养至汇合状态。细胞在N-NC凝胶上的粘附和增殖对粘土浓度(C(粘土))有很强的依赖性,培养细胞的数量在约C(粘土)=6×10(-2)摩尔(45.72克)/1升H(2)O时达到最大值。相反,在传统的化学交联PNIPA水凝胶上几乎不可能培养细胞,无论其交联剂浓度如何。从含水量、蛋白质吸附、表面平整度、脱水PNIPA链的疏水性以及剥离粘土上的阴离子电荷等方面讨论了细胞仅在具有特定网络结构和组成的N-NC凝胶表面培养的原因。最后发现,在N-NC凝胶表面培养的细胞无需胰蛋白酶处理,只需将温度降至20℃,就可以以细胞片的形式脱离。