Wang Lingyu, Yu Linfen, Grist Samantha, Cheung Karen C, Chen David D Y
Department of Chemistry, University of British Columbia, 2036 Main Mall, Vancouver, BC, V6T 1Z1 Canada.
Shenzhen Biorain Biotechnolgy Company, Suit A201, Shenwan 1 Road, Qianhai District, Shenzhen, 518100, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2017 Nov 15;1068-1069:105-111. doi: 10.1016/j.jchromb.2017.09.041. Epub 2017 Oct 12.
Cell culture systems based on polydimethylsiloxane (PDMS) microfluidic devices offer great flexibility because of their simple fabrication and adaptability. PDMS devices also make it straightforward to set up parallel experiments and can facilitate process automation, potentially speeding up the drug discovery process. However, cells grown in PDMS-based systems can develop in different ways to those grown with conventional culturing systems because of the differences in the containers' surfaces. Despite the growing number of studies on microfluidic cell culture devices, the differences in cellular behavior in PDMS-based devices and normal cell culture systems are poorly characterized. In this work, we investigated the proliferation and autophagy of MCF7 cells cultured in uncoated and Parylene-C coated PDMS wells. Using a quantitative method combining solid phase extraction and liquid chromatography mass spectrometry we developed, we showed that Tamoxifen uptake into the surfaces of uncoated PDMS wells can change the drug's effective concentration in the culture medium, affecting the results of Tamoxifen-induced autophagy and cytotoxicity assays. Such changes must be carefully analyzed before transferring in vitro experiments from a traditional culture environment to a PDMS-based microfluidic system. We also found that cells cultured in Parylene-C coated PDMS wells showed similar proliferation and drug response characteristics to cells cultured in standard polystyrene (PS) plates, indicating that Parylene-C deposition offers an easy way of limiting the uptake of small molecules into porous PDMS materials and significantly improves the performance of PDMS-based device for cell related research.
基于聚二甲基硅氧烷（PDMS）微流控装置的细胞培养系统具有很大的灵活性，因为其制造简单且适应性强。PDMS装置还使得设置平行实验变得直接，并且可以促进过程自动化，有可能加快药物发现过程。然而，由于容器表面的差异，在基于PDMS的系统中生长的细胞与在传统培养系统中生长的细胞相比，其发育方式可能不同。尽管对微流控细胞培养装置的研究越来越多，但基于PDMS的装置与正常细胞培养系统中细胞行为的差异仍未得到充分表征。在这项工作中，我们研究了在未涂层和聚对二甲苯-C涂层的PDMS孔中培养的MCF7细胞的增殖和自噬。使用我们开发的一种结合固相萃取和液相色谱质谱的定量方法，我们表明未涂层的PDMS孔表面对他莫昔芬的摄取会改变培养基中药物的有效浓度，从而影响他莫昔芬诱导的自噬和细胞毒性测定结果。在将体外实验从传统培养环境转移到基于PDMS的微流控系统之前，必须仔细分析这些变化。我们还发现，在聚对二甲苯-C涂层的PDMS孔中培养的细胞与在标准聚苯乙烯（PS）板中培养的细胞表现出相似的增殖和药物反应特性，这表明聚对二甲苯-C沉积提供了一种限制小分子进入多孔PDMS材料的简便方法，并显著提高了基于PDMS的细胞相关研究装置的性能。
