Department of Biomedical Engineering, University of California, Davis, Davis, CA 95616, USA.
Anal Bioanal Chem. 2012 Feb;402(5):1847-56. doi: 10.1007/s00216-011-5613-z. Epub 2011 Dec 29.
This paper describes a novel platform that utilizes micropatterning and electrochemistry to release cells-on-hydrogel microstructures from conductive indium tin oxide (ITO) substrates. In this approach, UV photopolymerization was employed to micropattern heparin-based hydrogels onto glass substrates containing ITO electrodes. ITO/glass substrates were first functionalized with acrylated silane to promote attachment of hydrogel structures. The surfaces containing hydrogel micropatterns were further functionalized with poly(ethylene glycol) thiol, rendering the regions around the hydrogel structures non-fouling to proteins and cells. After incubating surfaces with collagen (I), primary rat hepatocytes were shown to selectively attach on top of the hydrogel and not on surrounding glass/ITO regions. Electrical activation of specific ITO electrodes (-1.8 V vs. Ag/AgCl reference) was then used to release cells-on-hydrogel microstructures from the substrate. Immunostaining and reverse transcription polymerase chain reaction analysis of albumin, an important indicator of hepatic function, showed that the hepatocyte-on-hydrogel microstructures released from the surface maintained their function at levels similar to hepatocytes remaining on the culture substrate. In the future, switchable conductive substrates described here may be to collect cell samples at different time points and may also be used for harvesting cell-carrying vehicles for transplantation studies.
本文介绍了一种新颖的平台,利用微图案化和电化学将细胞-水凝胶微结构从导电铟锡氧化物(ITO)基底上释放出来。在这种方法中,采用紫外光聚合将基于肝素的水凝胶图案化到含有 ITO 电极的玻璃基底上。ITO/玻璃基底首先用丙烯酰基硅烷进行功能化,以促进水凝胶结构的附着。含有水凝胶微图案的表面进一步用聚乙二醇硫醇进行功能化,使水凝胶结构周围的区域对蛋白质和细胞无固着性。在用胶原蛋白(I)孵育表面后,发现原代大鼠肝细胞选择性地附着在水凝胶上,而不是周围的玻璃/ITO 区域。然后,通过对特定 ITO 电极进行电激活(相对于 Ag/AgCl 参比电极-1.8 V),将水凝胶上的细胞-微结构从基底上释放出来。对白蛋白(一种重要的肝功能指标)的免疫染色和逆转录聚合酶链反应分析表明,从表面释放的水凝胶上的肝细胞保留了与仍留在培养基底上的肝细胞相似的功能水平。在未来,这里描述的可切换导电基底可以用于在不同时间点收集细胞样本,也可以用于收获用于移植研究的载细胞载体。