Electrolytic valving isolation of cell co-culture microenvironment with controlled cell pairing ratios.
Electrolytic valving isolation of cell co-culture microenvironment with controlled cell pairing ratios.
复制标题
DOI：
10.1039/c4an01282h
复制
发表时间：
2014-12-21
期刊：
影响因子：
--
通讯作者：
Yoon E
中科院分区：
文献类型：
--
作者：
Chen YC;Ingram P;Yoon E
肿瘤间质相互作用是肿瘤发生的关键过程。传统的基于培养皿的共培养试验只是在同一个培养皿中混合两种细胞类型；因此，它们在控制细胞位置和精确跟踪异质细胞群体中的单细胞行为方面存在缺陷。微流控技术可以对微环境进行良好的时空控制，但这种控制通常是通过外部泵来实现的，这使得长期培养变得繁琐而笨重。在这项工作中，我们提出了一个细胞-细胞相互作用的微流控平台，该平台可以通过使用一种新的电解细胞隔离方案来精确控制共培养微环境，而无需使用任何阀门或气动泵。所提出的微流控平台还可以精确控制相互作用细胞的数量和配对比率，以模拟癌症生态位。超过80%的腔室捕获了所需数量的细胞。电池隔离的持续时间可以通过电解气泡的产生和去除来调节。在我们的平台上，我们验证了电解过程对细胞活力和增殖的影响可以忽略不计。据我们所知，这项工作是第一次尝试将电解气泡产生作为微流体中的细胞分离方法。为了证明其可行性，我们进行了前列腺癌（PC3）细胞和成肌细胞（C2C12）细胞之间的细胞间相互作用实验。初步结果表明，在细胞培养过程中利用电解进行微环境控制的潜力。比值控制的细胞-细胞相互作用实验也成功地表明，PC3与C2C12的细胞配对比例影响成肌细胞的增殖率，这是由于前列腺癌细胞分泌的生长因子增加。
Cancer-stromal interaction is a critical process in tumorigenesis. Conventional dish-based co-culture assays simply mix two cell types in the same dish; thus, they are deficient in controlling cell locations and precisely tracking single cell behavior from heterogeneous cell populations. Microfluidic technology can provide a good spatial temporal control of microenvironments, but the control has been typically realized by using external pumps, making long-term cultures cumbersome and bulky. In this work, we present a cell-cell interaction microfluidic platform that can accurately control co-culture microenvironment by using a novel electrolytic cell isolation scheme without using any valves or pneumatic pumps. The proposed microfluidic platform can also precisely control the number of interacting cells and pairing ratios to emulate cancer niches. More than 80% of the chambers captured the desired number of cells. The duration of cell isolation can be adjusted by electrolytic bubble generation and removal. We verified that electrolytic process has a negligible effect on cell viability and proliferation in our platform. To the best of our knowledge, this work is the first attempt to incorporate electrolytic bubble generation as a cell isolation method in microfluidics. For proof of feasibility, we performed cell-cell interaction assays between prostate cancer (PC3) cells and myoblast (C2C12) cells. The preliminary results demonstrated the potential of using electrolysis for micro-environmental control during cell culture. Also, the ratio controlled cell-cell interaction assays was successfully performed showing that the cell pairing ratios of PC3 to C2C12 affected the proliferation rate of myoblast cells due to increased secretion of growth factors from prostate cancer cells.