Paper-based microreactor integrating cell culture and subsequent immunoassay for the investigation of cellular phosphorylation.

Investigation of cellular phosphorylation and signaling pathway has recently gained much attention for the study of pathogenesis of cancer. Related conventional bioanalytical operations for this study including cell culture and Western blotting are time-consuming and labor-intensive. In this work, a paper-based microreactor has been developed to integrate cell culture and subsequent immunoassay on a single paper. The paper-based microreactor was a filter paper with an array of circular zones for running multiple cell cultures and subsequent immunoassays. Cancer cells were directly seeded in the circular zones without hydrogel encapsulation and cultured for 1 day. Subsequently, protein expressions including structural, functional, and phosphorylated proteins of the cells could be detected by their specific antibodies, respectively. Study of the activation level of phosphorylated Stat3 of liver cancer cells stimulated by IL-6 cytokine was demonstrated by the paper-based microreactor. This technique can highly reduce tedious bioanalytical operation and sample and reagent consumption. Also, the time required by the entire process can be shortened. This work provides a simple and rapid screening tool for the investigation of cellular phosphorylation and signaling pathway for understanding the pathogenesis of cancer. In addition, the operation of the paper-based microreactor is compatible to the molecular biological training, and therefore, it has the potential to be developed for routine protocol for various research areas in conventional bioanalytical laboratories.

[1]  Junfei Tian,et al.  Paper-based microfluidic devices by plasma treatment. , 2008, Analytical chemistry.

[2]  I. Papautsky,et al.  Optimization of a paper-based ELISA for a human performance biomarker. , 2013, Analytical chemistry.

[3]  M. Piquette-Miller,et al.  Influence of IL-6 on MDR and MRP-mediated multidrug resistance in human hepatoma cells. , 2001, Canadian journal of physiology and pharmacology.

[4]  Audrey K. Ellerbee,et al.  Quantifying colorimetric assays in paper-based microfluidic devices by measuring the transmission of light through paper. , 2009, Analytical chemistry.

[5]  Sindy K. Y. Tang,et al.  Multizone Paper Platform for 3D Cell Cultures , 2011, PloS one.

[6]  Hua Yu,et al.  Tumour immunology: Crosstalk between cancer and immune cells: role of STAT3 in the tumour microenvironment , 2007, Nature Reviews Immunology.

[7]  T. Golub,et al.  Epstein-Barr-Virus-Encoded LMP2A Induces Primary Epithelial Cell Migration and Invasion: Possible Role in Nasopharyngeal Carcinoma Metastasis , 2005, Journal of Virology.

[8]  Yan Liu,et al.  Inhibition of STAT3 Signaling Blocks the Anti-apoptotic Activity of IL-6 in Human Liver Cancer Cells* , 2010, The Journal of Biological Chemistry.

[9]  Sindy K. Y. Tang,et al.  Paper-supported 3D cell culture for tissue-based bioassays , 2009, Proceedings of the National Academy of Sciences.

[10]  S. Aguirre,et al.  Paper-based bioassays using gold nanoparticle colorimetric probes. , 2008, Analytical chemistry.

[11]  Jinghua Yu,et al.  Electrochemical immunoassay on a 3D microfluidic paper-based device. , 2012, Chemical Communications.

[12]  Kin Fong Lei,et al.  Microfluidic Systems for Diagnostic Applications , 2012, Journal of laboratory automation.

[13]  Fanyin Meng,et al.  Interleukin-6 and its receptor, key players in hepatobiliary inflammation and cancer. , 2012, Translational gastrointestinal cancer.

[14]  Anubhav Tripathi,et al.  Microfluidic reactors for diagnostics applications. , 2011, Annual review of biomedical engineering.

[15]  George M Whitesides,et al.  Polymer-based mesh as supports for multi-layered 3D cell culture and assays. , 2014, Biomaterials.

[16]  Emanuel Carrilho,et al.  Paper-based ELISA. , 2010, Angewandte Chemie.

[17]  G. Whitesides,et al.  Diagnostics for the developing world: microfluidic paper-based analytical devices. , 2010, Analytical chemistry.

[18]  Matthias W. Hentze,et al.  A bone morphogenetic protein (BMP)-responsive element in the hepcidin promoter controls HFE2-mediated hepatic hepcidin expression and its response to IL-6 in cultured cells , 2008, Journal of Molecular Medicine.

[19]  Qingliang Wang,et al.  The Expression of Functional Chemokine Receptor CXCR4 Is Associated with the Metastatic Potential of Human Nasopharyngeal Carcinoma , 2005, Clinical Cancer Research.

[20]  G. Whitesides,et al.  Understanding wax printing: a simple micropatterning process for paper-based microfluidics. , 2009, Analytical chemistry.

[21]  G. Whitesides.  The origins and the future of microfluidics , 2006, Nature.

[22]  Bingcheng Lin,et al.  Rapid prototyping of paper‐based microfluidics with wax for low‐cost, portable bioassay , 2009, Electrophoresis.

[23]  K. Vermeulen,et al.  The cell cycle: a review of regulation, deregulation and therapeutic targets in cancer , 2003, Cell proliferation.

[24]  Orawon Chailapakul,et al.  Electrochemical detection for paper-based microfluidics. , 2009, Analytical chemistry.

[25]  Jun Li,et al.  STAT3 activation in monocytes accelerates liver cancer progression , 2011, BMC Cancer.

[26]  George M Whitesides,et al.  Electrochemical sensing in paper-based microfluidic devices. , 2010, Lab on a chip.

[27]  George M Whitesides,et al.  FLASH: a rapid method for prototyping paper-based microfluidic devices. , 2008, Lab on a chip.

[28]  J. Rush,et al.  Immunoaffinity profiling of tyrosine phosphorylation in cancer cells , 2005, Nature Biotechnology.

[29]  C. Deng,et al.  Progenitor/stem cells give rise to liver cancer due to aberrant TGF-β and IL-6 signaling , 2008, Proceedings of the National Academy of Sciences.

[30]  Donald E Ingber,et al.  Platform for high-throughput testing of the effect of soluble compounds on 3D cell cultures. , 2013, Analytical chemistry.