Three Dimensional Microfluidic Cell Arrays for ex Vivo Drug Screening with Mimicked Vascular Flow

Currently, there are no reliable ex vivo models that predict anticancer drug responses in human tumors accurately. A comprehensive method of mimicking a 3D microenvironment to study effects of anticancer drugs on specific cancer types is essential. Here, we report the development of a three-dimensional microfluidic cell array (3D μFCA), which reconstructs a 3D tumor microenvironment with cancer cells and microvascular endothelial cells. To mimic the in vivo spatial relationship between microvessels and nonendothelial cells embedded in extracellular matrix, three polydimethylsiloxane (PDMS) layers were built into this array. The multilayer property of the device enabled the imitation of the drug delivery in a microtissue array with simulated blood circulation. This 3D μFCA system may provide better predictions of drug responses and identification of a suitable treatment for a specific patient if biopsy samples are used. To the pharmaceutical industry, the scaling-up of our 3D μFCA system may offer a novel high throughput screening tool.

[1]  Marilena Loizidou,et al.  3D tumour models: novel in vitro approaches to cancer studies , 2011, Journal of Cell Communication and Signaling.

[2]  John W Haycock,et al.  3D cell culture: a review of current approaches and techniques. , 2011, Methods in molecular biology.

[3]  A. Ellis Breast , 2002, BMJ : British Medical Journal.

[4]  G. Watanabe,et al.  Crosstalk to Stromal Fibroblasts Induces Resistance of Lung Cancer to Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitors , 2009, Clinical Cancer Research.

[5]  Luke P. Lee,et al.  Continuous perfusion microfluidic cell culture array for high-throughput cell-based assays. , 2005, Biotechnology and bioengineering.

[6]  R. Rosenfeld Nature , 2009, Otolaryngology--head and neck surgery : official journal of American Academy of Otolaryngology-Head and Neck Surgery.

[7]  John Greenman,et al.  A Microfluidic System for Testing the Responses of Head and Neck Squamous Cell Carcinoma Tissue Biopsies to Treatment with Chemotherapy Drugs , 2011, Annals of Biomedical Engineering.

[8]  A. Jayaraman,et al.  Dynamic gene expression profiling using a microfabricated living cell array. , 2004, Analytical chemistry.

[9]  Hanry Yu,et al.  A novel 3D mammalian cell perfusion-culture system in microfluidic channels. , 2007, Lab on a chip.

[10]  D Liberati,et al.  Forecasting the growth of multicell tumour spheroids: implications for the dynamic growth of solid tumours , 2000, Cell proliferation.

[11]  John Greenman,et al.  Development of a microfluidic device for the maintenance and interrogation of viable tissue biopsies. , 2008, Lab on a chip.

[12]  Luke P. Lee,et al.  Nanoliter scale microbioreactor array for quantitative cell biology , 2006, Biotechnology and bioengineering.

[13]  R. Kamm,et al.  Three-dimensional microfluidic model for tumor cell intravasation and endothelial barrier function , 2012, Proceedings of the National Academy of Sciences.

[14]  John Greenman,et al.  Microfluidic perfusion system for maintaining viable heart tissue with real-time electrochemical monitoring of reactive oxygen species. , 2010, Lab on a chip.

[15]  Ciprian Iliescu,et al.  Exploitation of physical and chemical constraints for three-dimensional microtissue construction in microfluidics. , 2011, Biomicrofluidics.

[16]  D. Sabatini,et al.  Microarrays of cells expressing defined cDNAs , 2001, Nature.

[17]  S. Quake,et al.  Versatile, fully automated, microfluidic cell culture system. , 2007, Analytical chemistry.

[18]  J. Voldman,et al.  Microfluidic arrays for logarithmically perfused embryonic stem cell culture. , 2006, Lab on a chip.

[19]  C. Redfern,et al.  In vivo-like drug responses of human tumors growing in three-dimensional gel-supported primary culture. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[20]  M. Koutsilieris,et al.  Three-dimensional culture system as a model for studying cancer cell invasion capacity and anticancer drug sensitivity. , 2004, Anticancer research.

[21]  E. Verpoorte,et al.  Hydrogel embedding of precision‐cut liver slices in a microfluidic device improves drug metabolic activity , 2011, Biotechnology and bioengineering.

[22]  Shuichi Takayama,et al.  Microfluidic Endothelium for Studying the Intravascular Adhesion of Metastatic Breast Cancer Cells , 2009, PloS one.

[23]  宁北芳,et al.  疟原虫var基因转换速率变化导致抗原变异[英]/Paul H, Robert P, Christodoulou Z, et al//Proc Natl Acad Sci U S A , 2005 .

[24]  E. Verpoorte,et al.  An alternative approach based on microfluidics to study drug metabolism and toxicity using liver and intestinal tissue , 2010 .

[25]  Roger D Kamm,et al.  Screening therapeutic EMT blocking agents in a three-dimensional microenvironment. , 2013, Integrative biology : quantitative biosciences from nano to macro.

[26]  Chu Zhang,et al.  Hyaluronic acid-based hydrogels as 3D matrices for in vitro evaluation of chemotherapeutic drugs using poorly adherent prostate cancer cells. , 2009, Biomaterials.

[27]  E. Golemis,et al.  Fibroblast-derived 3D matrix differentially regulates the growth and drug-responsiveness of human cancer cells. , 2008, Matrix biology : journal of the International Society for Matrix Biology.

[28]  Mehmet Toner,et al.  A high-throughput microfluidic real-time gene expression living cell array. , 2007, Lab on a chip.

[29]  J. Tarbell,et al.  Effect of the glycocalyx layer on transmission of interstitial flow shear stress to embedded cells , 2012, Biomechanics and Modeling in Mechanobiology.

[30]  Roman Rouzier,et al.  Change in tumor cellularity of breast carcinoma after neoadjuvant chemotherapy as a variable in the pathologic assessment of response , 2004, Cancer.

[31]  R. Hoffman,et al.  In vivo-like growth of human tumors in vitro. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[32]  Hans G Othmer,et al.  The role of the microenvironment in tumor growth and invasion. , 2011, Progress in biophysics and molecular biology.

[33]  B. Sipos,et al.  Tumor Stroma Interactions Induce Chemoresistance in Pancreatic Ductal Carcinoma Cells Involving Increased Secretion and Paracrine Effects of Nitric Oxide and Interleukin-1β , 2004, Cancer Research.

[34]  B. Lin,et al.  Cell-based high content screening using an integrated microfluidic device. , 2007, Lab on a chip.

[35]  Robert L Sah,et al.  Probing the role of multicellular organization in three-dimensional microenvironments , 2006, Nature Methods.

[36]  Teruo Fujii,et al.  Bile canaliculi formation by aligning rat primary hepatocytes in a microfluidic device. , 2011, Biomicrofluidics.

[37]  S. Bhatia,et al.  An extracellular matrix microarray for probing cellular differentiation , 2005, Nature Methods.

[38]  J. Pollard,et al.  Microenvironmental regulation of metastasis , 2009, Nature Reviews Cancer.

[39]  D. Ingber,et al.  Reconstituting Organ-Level Lung Functions on a Chip , 2010, Science.

[40]  V. D’Andrea,et al.  Distribution of acetylcholinesterase and cholineacetyl‐transferase activities in the human pulmonary vessels of younger and older adults , 2005 .

[41]  D. Yamamoto,et al.  Influence of cellularity in human breast carcinoma. , 2004, Breast.

[42]  Sophie Lelièvre,et al.  beta4 integrin-dependent formation of polarized three-dimensional architecture confers resistance to apoptosis in normal and malignant mammary epithelium. , 2002, Cancer cell.

[43]  R K Jain,et al.  Vascular permeability in a human tumor xenograft: molecular size dependence and cutoff size. , 1995, Cancer research.

[44]  Peter Vaupel,et al.  Tumor microenvironmental physiology and its implications for radiation oncology. , 2004, Seminars in radiation oncology.

[45]  Ivan Martin,et al.  New dimensions in tumor immunology: what does 3D culture reveal? , 2008, Trends in molecular medicine.

[46]  Anne E Carpenter,et al.  Microarrays of lentiviruses for gene function screens in immortalized and primary cells , 2006, Nature Methods.

[47]  D. Beebe,et al.  Microenvironment design considerations for cellular scale studies. , 2004, Lab on a chip.

[48]  Z. Hall Cancer , 1906, The Hospital.

[49]  L. Holmberg,et al.  Quantification of Normal Cell Fraction and Copy Number Neutral LOH in Clinical Lung Cancer Samples Using SNP Array Data , 2009, PloS one.