Transparent polymeric cell culture chip with integrated temperature control and uniform media perfusion
Transparent polymeric cell culture chip with integrated temperature control and uniform media perfusion
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DOI：
10.2144/000112122
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发表时间：
2006-03-01
期刊：
影响因子：
2.7
通讯作者：
Dufva, M
中科院分区：
文献类型：
--
作者：
Petronis, S;Stangegaard, M;Dufva, M
现代微加工和微流体技术为设计和制造用于活细胞在线监测的微型化细胞培养系统提供了新的机会。我们使用激光微加工和热粘合来制造光学透明的、低成本的聚合物芯片，用于在受控条件下长期在线观察细胞培养。该芯片结合了微流体流动均衡系统，确保细胞培养基在整个细胞培养室中的均匀灌注。集成的氧化铟锡加热器和微型温度探头连接到电子反馈系统，创造了稳定和空间均匀的热条件，对芯片的光学透明度的干扰最小。芯片的流体和热性能进行了验证，通过有限元建模和波动的环境温度条件下的操作测试。HeLa细胞在细胞培养芯片内培养长达2周，并使用延时视频记录显微镜装置进行监测。在前10-20小时（滞后期）期间观察到细胞附着和铺展。大约20小时后，细胞生长呈指数特征，估计倍增时间约为32小时，这与在CO2培养箱中的标准细胞培养瓶中生长的细胞观察到的倍增时间相同。
Modern microfabrication and microfluidic technologies offer new opportunities in the design and fabrication of miniaturized cell culture systems for online monitoring of living cells. We used laser micromachining and thermal bonding to fabricate an optically transparent, low-cost polymeric chip for long-term online cell culture observation under controlled conditions. The chip incorporated a microfluidic flow equalization system, assuring uniform perfusion of the cell culture media throughout the cell culture chamber. The integrated indium-tin-oxide heater and miniature temperature probe linked to an electronic feedback system created steady and spatially uniform thermal conditions with minimal interference to the optical transparency of the chip. The fluidic and thermal performance of the chip was verified by finite element modeling and by operation tests under fluctuating ambient temperature conditions. HeLa cells were cultured for up to 2 weeks within the cell culture chip and monitored using a time-lapse video recording microscopy setup. Cell attachment and spreading was observed during the first 10-20 h (lag phase). After approximately 20 h, cell growth gained exponential character with an estimated doubling time of about 32 h, which is identical to the observed doubling time of cells grown in standard cell culture flasks in a CO2 incubator.