Hung Paul J, Lee Philip J, Sabounchi Poorya, Lin Robert, Lee Luke P
Berkeley Sensor & Actuator Center, Department of Bioengineering, University of California, 485 Evans Hall, Berkeley, CA 94720, USA.
Biotechnol Bioeng. 2005 Jan 5;89(1):1-8. doi: 10.1002/bit.20289.
We present for the first time a microfluidic cell culture array for long-term cellular monitoring. The 10 x 10 array could potentially assay 100 different cell-based experiments in parallel. The device was designed to integrate the processes used in typical cell culture experiments on a single self-contained microfluidic system. Major functions include repeated cell growth/passage cycles, reagent introduction, and real-time optical analysis. The single unit of the array consists of a circular microfluidic chamber, multiple narrow perfusion channels surrounding the main chamber, and four ports for fluidic access. Human carcinoma (HeLa) cells were cultured inside the device with continuous perfusion of medium at 37 degrees C. The observed doubling time was 1.4 +/- 0.1 days with a peak cell density of approximately 2.5*10(5) cells/cm(2). Cell assay was demonstrated by monitoring the fluorescence localization of calcein AM from 1 min to 10 days after reagent introduction. Confluent cell cultures were passaged within the microfluidic chambers using trypsin and successfully regrown, suggesting a stable culture environment suitable for continuous operation. The cell culture array could offer a platform for a wide range of assays with applications in drug screening, bioinformatics, and quantitative cell biology.
我们首次展示了一种用于长期细胞监测的微流控细胞培养阵列。这个10×10的阵列有可能同时检测100个不同的基于细胞的实验。该设备旨在将典型细胞培养实验中使用的过程集成到一个独立的微流控系统中。主要功能包括重复的细胞生长/传代循环、试剂引入和实时光学分析。阵列的单个单元由一个圆形微流控腔室、围绕主腔室的多个狭窄灌注通道以及四个流体接入端口组成。人癌细胞（HeLa）在该设备内于37摄氏度下持续灌注培养基进行培养。观察到的倍增时间为1.4±0.1天，峰值细胞密度约为2.5×10⁵个细胞/cm²。通过监测试剂引入后1分钟至10天内钙黄绿素AM的荧光定位来进行细胞检测。使用胰蛋白酶在微流控腔内对汇合的细胞培养物进行传代，并成功重新生长，这表明存在适合连续操作的稳定培养环境。该细胞培养阵列可为药物筛选、生物信息学和定量细胞生物学等广泛的检测提供一个平台。
