Biological implications of polydimethylsiloxane-based microfluidic cell culture.
Biological implications of polydimethylsiloxane-based microfluidic cell culture.
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DOI：
10.1039/b903043c
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发表时间：
2009-08-07
期刊：
影响因子：
6.1
通讯作者：
Beebe DJ
中科院分区：
文献类型：
--
作者：
Regehr KJ;Domenech M;Koepsel JT;Carver KC;Ellison-Zelski SJ;Murphy WL;Schuler LA;Alarid ET;Beebe DJ
聚二甲基硅氧烷（PDMS）由于其简单的制造工艺和材料属性（如透气性、光学透明度和柔性）已成为微流体领域的主要产品。随着微流体系统被用于生物学问题，并越来越多地用作细胞培养平台，PDMS的材料特性必须在生物学背景下考虑。在这项研究中，PDMS的两个属性：未固化的低聚物从聚合物网络到微通道介质的浸出，和小的，疏水分子（即雌激素）从含血清的介质到聚合物块的吸收。未固化的PDMS低聚物可通过MALDI-MS在乙醇中对PDMS装置进行索氏提取之前和之后在微通道介质中检测到。此外，在PDMS微通道中培养24小时的NMuMG细胞的质膜中鉴定了PDMS寡聚体。在提取的微通道中培养的细胞也含有可检测量的未固化的PDMS。结果表明，直接接种在PDMS插入物上的MCF-7细胞对亲水性催乳素有反应，但对疏水性雌激素无反应，反映了其对吸收小的疏水分子的特异性;漂浮在威尔斯孔中的PDMS的存在以血清依赖性方式显著降低了细胞对雌激素的反应。通过ELISA对雌激素进行定量显示，微通道雌激素以约9：1的比例迅速分配到周围的PDMS中。预处理，如阻断血清或预吸收雌激素24小时不影响雌激素从PDMS为基础的微通道损失。这些发现强调了在确定生物实验的适当环境时仔细考虑培养系统特性的重要性。
Polydimethylsiloxane (PDMS) has become a staple of the microfluidics community by virtue of its simple fabrication process and material attributes, such as gas permeability, optical transparency, and flexibility. As microfluidic systems are put toward biological problems and increasingly utilized as cell culture platforms, the material properties of PDMS must be considered in a biological context. Two properties of PDMS were addressed in this study: the leaching of uncured oligomers from the polymer network into microchannel media, and the absorption of small, hydrophobic molecules (i.e. estrogen) from serum-containing media into the polymer bulk. Uncured PDMS oligomers were detectable via MALDI-MS in microchannel media both before and after Soxhlet extraction of PDMS devices in ethanol. Additionally, PDMS oligomers were identified in the plasma membranes of NMuMG cells cultured in PDMS microchannels for 24 hours. Cells cultured in extracted microchannels also contained a detectable amount of uncured PDMS. It was shown that MCF-7 cells seeded directly on PDMS inserts were responsive to hydrophilic prolactin but not hydrophobic estrogen, reflecting its specificity for absorbing small, hydrophobic molecules; and the presence of PDMS floating in wells significantly reduced cellular response to estrogen in a serum-dependent manner. Quantification of estrogen via ELISA revealed that microchannel estrogen partitioned rapidly into the surrounding PDMS to a ratio of approximately 9:1. Pretreatments such as blocking with serum or pre-absorbing estrogen for 24 hours did not affect estrogen loss from PDMS-based microchannels. These findings highlight the importance of careful consideration of culture system properties when determining an appropriate environment for biological experiments.
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影响因子：
6.1
作者：
Millet, Larry J.;Stewart, Matthew E.;Gillette, Martha U.
通讯作者：
Gillette, Martha U.
影响因子：
6.1
作者：
Abate, Adam R.;Lee, Daeyeon;Weitz, David A.
通讯作者：
Weitz, David A.
影响因子：
2.7
作者：
Jo, BH;Van Lerberghe, LM;Beebe, DJ
通讯作者：
Beebe, DJ
影响因子：
3.9
作者：
Jiang, XY;Takayama, S;Whitesides, GM
通讯作者：
Whitesides, GM
影响因子：
3
作者：
TOTH, A;BERTOTI, I;SZAPLONCZAY, P
通讯作者：
SZAPLONCZAY, P