Stable immobilization of rat hepatocytes as hemispheroids onto collagen-conjugated poly-dimethylsiloxane (PDMS) surfaces: Importance of direct oxygenation through PDMS for both formation and function
Stable immobilization of rat hepatocytes as hemispheroids onto collagen-conjugated poly-dimethylsiloxane (PDMS) surfaces: Importance of direct oxygenation through PDMS for both formation and function
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DOI：
10.1002/bit.21690
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发表时间：
2008-04-15
影响因子：
3.8
通讯作者：
Sakai, Yasuyuki
中科院分区：
文献类型：
--
作者：
Nishikawa, Masaki;Yamamoto, Takatoki;Sakai, Yasuyuki
高透氧材料聚二甲基硅氧烷（PDMS）具有应用于细胞培养微型装置的潜力，“但细胞从PDMS上脱落一直是一个主要问题。在这项研究中，我们证明，胶原共价固定的PDMS和足够的氧气供应的组合，使建立一个稳定的，附着的球体（半球形）培养大鼠肝细胞。底部PDMS表面首先用氧等离子体处理，然后与氨基硅烷偶联，随后与pbotoreactive交联剂偶联，最后与胶原蛋白溶液反应。X射线光电子能谱（XPS）和接触角的测量表明，共价固定的胶原蛋白的表面上发生的交联剂已被引入。在胶原结合的PDMS表面上，大鼠肝细胞将自己组织成半球形，并保持活力和显着高的白蛋白产生至少2周的培养。相比之下，其他类型的PDMS表面上的肝细胞形成悬浮的球状体，其具有低白蛋白产生。此外，我们发现，通过底部PDMS表面阻断氧气供应抑制半球体的形成和肝细胞功能的增强。这些结果表明，适当的表面改性的PDMS是一个有前途的方法对肝组织微器件的发展。
The highly oxygen-permeable material, polydimethylsiloxane (PDMS), has the potential to be applied to cell culture microdevices,"but cell detachment from PDMS has been a major problem. In this study, we demonstrate that a combination of collagen covalently immobilized PDMS and an adequate oxygen supply enables the establishment of a stable, attached spheroid (hemispheroid) culture of rat hepatocytes. The bottom PDMS surfaces were first treated with oxygen plasma, then coupled with aminosilane followed by a pbotoreactive crosslinker, and they were finally reacted with a collagen solution. X-ray photoelectron spectroscopy (XPS) and contact angle measurements showed that the covalent immobilization of collagen on the surface occurred only where the crosslinker had been introduced. On the collagen-conjugated PDMS surface, rat hepatocytes organized themselves into hemispheroids and maintained the viability and a remarkably high albumin production at least for 2 weeks of culture. In contrast, hepatocytes on the other types of PDMS surfaces formed suspended spheroids that had low albumin production. In addition, we showed that blocking the oxygen supply through the bottom PDMS surface inhibited the formation of hemispheroids and the augmentation of hepatocellular function. These results show that appropriate surface modification of PDMS is a promising approach towards the development of liver tissue microdevices.