A novel 3-D bio-microfluidic system mimicking in vivo heterogeneous tumour microstructures reveals complex tumour-stroma interactions.

A 3-D microfluidic system consisting of microchamber arrays embedded in a collagen hydrogel with tuneable biochemical gradients that mimics the tumour microenvironment of mammary glands was constructed for the investigation on the interactions between invasive breast cancer cells and stromal cells. The hollow microchambers in collagen provide a very similar 3-D environment to that in vivo that regulates collective cellular dynamics and behaviour, while the microfluidic channels surrounding the collagen microchamber arrays allow one to impose complex concentration gradients of specific biological molecules or drugs. We found that breast epithelial cells (MCF-10A) seeded in the microchambers formed lumen-like structures similar to those in epithelial layers. When MCF-10A cells were co-cultured with invasive breast cancer cells (MDA-MB-231), the formation of lumen-like structures in the microchambers was inhibited, indicating the capability of cancer cells to disrupt the structures formed by surrounding cells for further invasion and metastasis. Subsequent mechanism studies showed that down regulation of E-cad expression due to MMPs produced by the cancer cells plays a dominant role in determining the cellular behaviour. Our microfluidic system offers a robust platform for high throughput studies that aim to understand combinatorial effects of multiple biochemical and microenvironmental factors.

[1]  Wenqi Hu,et al.  An opto-thermocapillary cell micromanipulator. , 2013, Lab on a chip.

[2]  M. Poo,et al.  Endothelial cell polarization and chemotaxis in a microfluidic device. , 2008, Lab on a chip.

[3]  Denis Wirtz,et al.  The physics of cancer: the role of physical interactions and mechanical forces in metastasis , 2011, Nature Reviews Cancer.

[4]  M. Shen.  Cancer: The complex seeds of metastasis , 2015, Nature.

[5]  Mehdi Nikkhah,et al.  MCF10A and MDA-MB-231 human breast basal epithelial cell co-culture in silicon micro-arrays. , 2011, Biomaterials.

[6]  B. Karlan,et al.  Evaluation of MCF10A as a Reliable Model for Normal Human Mammary Epithelial Cells , 2015, PloS one.

[7]  Samuel K Sia,et al.  In situ collagen assembly for integrating microfabricated three-dimensional cell-seeded matrices. , 2008, Nature materials.

[8]  L. Behrend,et al.  Manganese Superoxide Dismutase Induces p53-Dependent Senescence in Colorectal Cancer Cells , 2005, Molecular and Cellular Biology.

[9]  Kevin Barraclough,et al.  I and i , 2001, BMJ : British Medical Journal.

[10]  Guillaume Lambert,et al.  Cell motility and drug gradients in the emergence of resistance to chemotherapy , 2013, Proceedings of the National Academy of Sciences.

[11]  A. Khademhosseini,et al.  BIOMIMETIC GRADIENT HYDROGELS FOR TISSUE ENGINEERING. , 2010, The Canadian journal of chemical engineering.

[12]  X. Guan,et al.  Cancer metastases: challenges and opportunities , 2015, Acta pharmaceutica Sinica. B.

[13]  E. Pietras,et al.  JCB: Review , 2011 .

[14]  Moumita Das,et al.  Dynamic self-organization of microwell-aggregated cellular mixtures. , 2016, Soft matter.

[15]  W. Marsden.  I and J , 2012 .

[16]  Denis Wirtz,et al.  Mismatch in mechanical and adhesive properties induces pulsating cancer cell migration in epithelial monolayer. , 2012, Biophysical journal.

[17]  Aaas News,et al.  Book Reviews , 1893, Buffalo Medical and Surgical Journal.

[18]  M. Bissell,et al.  Three-dimensional lithographically defined organotypic tissue arrays for quantitative analysis of morphogenesis and neoplastic progression , 2008, Nature Protocols.

[19]  W. Fan,et al.  Different administration strategies with paclitaxel induce distinct phenotypes of multidrug resistance in breast cancer cells. , 2013, Cancer letters.

[20]  JONG BIN Kim,et al.  Three-dimensional tissue culture models in cancer biology. , 2005, Seminars in cancer biology.

[21]  R. Stephenson.  A and V , 1962, The British journal of ophthalmology.

[22]  Wenqi Hu,et al.  Hydrogel microrobots actuated by optically generated vapour bubbles. , 2012, Lab on a chip.

[23]  Joe Tien,et al.  Molding of three-dimensional microstructures of gels. , 2003, Journal of the American Chemical Society.

[24]  Bingcheng Lin,et al.  Carcinoma-associated fibroblasts promoted tumor spheroid invasion on a microfluidic 3D co-culture device. , 2010, Lab on a chip.

[25]  Martin Clynes,et al.  In vitro Development of Chemotherapy and Targeted Therapy Drug-Resistant Cancer Cell Lines: A Practical Guide with Case Studies , 2014, Front. Oncol..

[26]  Mingming Wu,et al.  Cooperative Roles of SDF-1α and EGF Gradients on Tumor Cell Migration Revealed by a Robust 3D Microfluidic Model , 2013, PloS one.

[27]  P. Drew,et al.  The analysis of doxorubicin resistance in human breast cancer cells using antibody microarrays , 2006, Molecular Cancer Therapeutics.

[28]  Shikuan Yang,et al.  Theory and experiment on particle trapping and manipulation via optothermally generated bubbles. , 2014, Lab on a chip.

[29]  Yunfeng Zhao,et al.  Manganese superoxide dismutase in cancer prevention. , 2014, Antioxidants & redox signaling.

[30]  Valerie M. Weaver,et al.  A tense situation: forcing tumour progression , 2009, Nature Reviews Cancer.

[31]  M. Dewhirst,et al.  Cellular redox modulator, ortho Mn(III) meso-tetrakis(N-n-hexylpyridinium-2-yl)porphyrin, MnTnHex-2-PyP(5+) in the treatment of brain tumors. , 2011, Anti-cancer agents in medicinal chemistry.

[32]  D. Albanes,et al.  Manganese superoxide dismutase (MnSOD) polymorphism, α-tocopherol supplementation and prostate cancer risk in the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study (Finland) , 2003, Cancer Causes & Control.

[33]  Dietmar W Hutmacher,et al.  Biomaterials offer cancer research the third dimension. , 2010, Nature materials.

[34]  Andrew J Ewald,et al.  ECM microenvironment regulates collective migration and local dissemination in normal and malignant mammary epithelium , 2012, Proceedings of the National Academy of Sciences.

[35]  M. Gottesman,et al.  Beyond 3D culture models of cancer , 2015, Science Translational Medicine.

[36]  B. Mian,et al.  Manganese superoxide dismutase enhances the invasive and migratory activity of tumor cells. , 2007, Cancer research.

[37]  A. Rajasekaran,et al.  Carcinoma cells induce lumen filling and EMT in epithelial cells through soluble E-cadherin-mediated activation of EGFR , 2015, Journal of Cell Science.

[38]  Pei-Hsun Wu,et al.  Normal mammary epithelial cells promote carcinoma basement membrane invasion by inducing microtubule-rich protrusions , 2015, Oncotarget.

[39]  Raquel Perez-Castillejos,et al.  Partitioning microfluidic channels with hydrogel to construct tunable 3-D cellular microenvironments. , 2008, Biomaterials.