Cultivation and recovery of vascular endothelial cells in microchannels of a separable micro-chemical chip.
Cultivation and recovery of vascular endothelial cells in microchannels of a separable micro-chemical chip.
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DOI：
10.1016/j.biomaterials.2010.12.012
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发表时间：
2011-04
期刊：
影响因子：
14
通讯作者：
Tadahiro Yamashita;Yo Tanaka;N. Idota;Kae Sato;K. Mawatari;T. Kitamori
中科院分区：
文献类型：
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作者：
Tadahiro Yamashita;Yo Tanaka;N. Idota;Kae Sato;K. Mawatari;T. Kitamori
最近开发了各种微细胞培养系统。然而，从微通道中恢复培养细胞是极其困难的，因为微芯片的上下底物是永久结合的。因此，我们开发了一种细胞培养和恢复系统，该系统使用具有可逆结合的可分离微芯片，允许封闭通道和开放通道之间的分离。为了实现这一系统，必须克服与微流体控制相关的两个问题——防止泄漏和从基质中无创地恢复培养细胞。在本研究中，我们采用表面化学改性的方法来解决这两个问题。首先，利用十八烷基三甲氧基硅烷（ODTMS）控制液/气相界面处的拉普拉斯压力，使其指向微通道，从而抑制两个衬底之间微小间隙的泄漏。其次，利用热响应性聚合物聚n -异丙基丙烯酰胺（PNIPAAm）在odtms修饰的微通道表面进行光聚合。PNIPAAm底物是众所周知的通过改变温度来控制细胞粘附/脱离。最后，对ODTMS-和pnipaam修饰的可分离微芯片进行图图化处理，并在无渗漏的微通道中培养人动脉内皮细胞（HAECs）。培养96 h后，通过降低温度将haec与微芯片分离，然后从微通道中回收。这项研究首次证明了在微通道中培养活细胞的恢复，并可能作为血管组织工程的基础技术。
Various micro cell culture systems have recently been developed. However, it is extremely difficult to recover cultured cells from a microchannel because the upper and lower substrates of a microchip are permanently combined. Therefore, we developed a cell culture and recovery system that uses a separable microchip with reversible combining that allows separation between closed and open channels. To realize this system, two problems related to microfluidic control—prevention of leakage and non-invasive recovery of cultured cells from the substrate—must be overcome. In the present study, we used surface chemistry modification to solve both problems. First, octadecyltrimethoxysilane (ODTMS) was utilized to control the Laplace pressure at the liquid/vapor phase interface, such that it was directed toward the microchannels, which suppressed leakage from the slight gap between two substrates. Second, a thermoresponsive polymer poly(N-isopropyl acrylamide) (PNIPAAm) was used to coat the surface of the ODTMS-modified microchannel by UV-mediated photopolymerization. PNIPAAm substrates are well known for controlled cell adhesion/detachment by alteration of temperature. Finally, the ODTMS- and PNIPAAm-modified separable microchips were subjected to patterning, and human arterial endothelial cells (HAECs) were cultured in the resulting microchannels with no leakage. After 96 h of the culture, the HAECs were detached from the microchips by decreasing the temperature and were then recovered from the microchannels. This study is the first to demonstrate the recovery of living cells cultured in a microchannel, and may be useful as a fundamental technique for vascular tissue engineering.