Microdroplet chain array for cell migration assays.

Establishing cell migration assays in multiple different microenvironments is important in the study of tissue repair and regeneration, cancer progression, atherosclerosis, and arthritis. In this work, we developed a miniaturized and massive parallel microfluidic platform for multiple cell migration assays combining the traditional membrane-based cell migration technique and the droplet-based microfluidic technique. Nanoliter-scale droplets are flexibly assembled as building blocks based on a porous membrane to form microdroplet chains with diverse configurations for different assay modes. Multiple operations including in-droplet 2D/3D cell culture, cell co-culture and cell migration induced by a chemoattractant concentration gradient in droplet chains could be flexibly performed with reagent consumption in the nanoliter range for each assay and an assay scale-up to 81 assays in parallel in one microchip. We have applied the present platform to multiple modes of cell migration assays including the accurate cell migration assay, competitive cell migration assay, biomimetic chemotaxis assay, and multifactor cell migration assay based on the organ-on-a-chip concept, for demonstrating its versatility, applicability, and potential in cell migration-related research.

[1]  S. Boyden.  THE CHEMOTACTIC EFFECT OF MIXTURES OF ANTIBODY AND ANTIGEN ON POLYMORPHONUCLEAR LEUCOCYTES , 1962, The Journal of experimental medicine.

[2]  D. Lauffenburger,et al.  Cell Migration: A Physically Integrated Molecular Process , 1996, Cell.

[3]  A. Mikos,et al.  Inhibition of smooth muscle cell growth in vitro by an antisense oligodeoxynucleotide released from poly(DL-lactic-co-glycolic acid) microparticles. , 1997, Journal of biomedical materials research.

[4]  M. Jordan,et al.  Microtubules and actin filaments: dynamic targets for cancer chemotherapy. , 1998, Current opinion in cell biology.

[5]  S. Quake,et al.  From micro- to nanofabrication with soft materials. , 2000, Science.

[6]  G. Whitesides,et al.  Neutrophil chemotaxis in linear and complex gradients of interleukin-8 formed in a microfabricated device , 2002, Nature Biotechnology.

[7]  G. Borisy,et al.  Cell Migration: Integrating Signals from Front to Back , 2003, Science.

[8]  Ali Khademhosseini,et al.  Fabrication of gradient hydrogels using a microfluidics/photopolymerization process. , 2004, Langmuir : the ACS journal of surfaces and colloids.

[9]  Yumiko Sakai,et al.  A novel in vitro system, the integrated discrete multiple organ cell culture (IdMOC) system, for the evaluation of human drug toxicity: comparative cytotoxicity of tamoxifen towards normal human cells from five major organs and MCF-7 adenocarcinoma breast cancer cells. , 2004, Chemico-biological interactions.

[10]  Lars Nielsen,et al.  Hanging-drop multicellular spheroids as a model of tumour angiogenesis , 2004, Angiogenesis.

[11]  J. Peterse,et al.  Breast cancer metastasis: markers and models , 2005, Nature Reviews Cancer.

[12]  David J Beebe,et al.  Characterization of a membrane-based gradient generator for use in cell-signaling studies. , 2006, Lab on a chip.

[13]  A. Albini,et al.  The chemoinvasion assay: a method to assess tumor and endothelial cell invasion and its modulation , 2007, Nature Protocols.

[14]  A. Lee,et al.  Engineering microscale cellular niches for three-dimensional multicellular co-cultures. , 2009, Lab on a chip.

[15]  Jong Hwan Sung,et al.  A micro cell culture analog (microCCA) with 3-D hydrogel culture of multiple cell lines to assess metabolism-dependent cytotoxicity of anti-cancer drugs. , 2009, Lab on a chip.

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

[17]  D. Beebe,et al.  Fundamentals of microfluidic cell culture in controlled microenvironments. , 2010, Chemical Society reviews.

[18]  Niraj K Inamdar,et al.  Microfluidic cell culture models for tissue engineering. , 2011, Current opinion in biotechnology.

[19]  Chang Lu,et al.  Chemical transfection of cells in picoliter aqueous droplets in fluorocarbon oil. , 2011, Analytical chemistry.

[20]  Anna Grazia Monteduro,et al.  Automatic transwell assay by an EIS cell chip to monitor cell migration. , 2011, Lab on a chip.

[21]  N. Melosh,et al.  Rapid spatial and temporal controlled signal delivery over large cell culture areas. , 2011, Lab on a chip.

[22]  Shawn M. Gomez,et al.  Arp2/3 Is Critical for Lamellipodia and Response to Extracellular Matrix Cues but Is Dispensable for Chemotaxis , 2012, Cell.

[23]  R. Kamm,et al.  Microfluidic assay for simultaneous culture of multiple cell types on surfaces or within hydrogels , 2012, Nature Protocols.

[24]  Jing Liu,et al.  Soft fibrin gels promote selection and growth of tumourigenic cells , 2012, Nature Materials.

[25]  D. Beebe,et al.  Microfluidic kit-on-a-lid: a versatile platform for neutrophil chemotaxis assays. , 2012, Blood.

[26]  Hon Fai Chan,et al.  Rapid formation of multicellular spheroids in double-emulsion droplets with controllable microenvironment , 2013, Scientific Reports.

[27]  Savas Tasoglu,et al.  Manipulating biological agents and cells in micro-scale volumes for applications in medicine. , 2013, Chemical Society reviews.

[28]  Ying Zhu,et al.  Cell-based drug combination screening with a microfluidic droplet array system. , 2013, Analytical chemistry.

[29]  Uwe Marx,et al.  Skin and hair on-a-chip: in vitro skin models versus ex vivo tissue maintenance with dynamic perfusion. , 2013, Lab on a chip.

[30]  Q. Fang,et al.  Sequential operation droplet array: an automated microfluidic platform for picoliter-scale liquid handling, analysis, and screening. , 2013, Analytical chemistry.

[31]  Timothy J. Mitchison,et al.  Biased migration of confined neutrophil-like cells in asymmetric hydraulic environments , 2013, Proceedings of the National Academy of Sciences.

[32]  Xavier Trepat,et al.  Propulsion and navigation within the advancing monolayer sheet , 2013, Nature materials.

[33]  D J Beebe,et al.  Gradient generation platforms: new directions for an established microfluidic technology. , 2014, Lab on a chip.

[34]  Matteo Moretti,et al.  In vitro models of the metastatic cascade: from local invasion to extravasation. , 2014, Drug discovery today.

[35]  R. Huang,et al.  Modeling of cancer metastasis and drug resistance via biomimetic nano-cilia and microfluidics. , 2014, Biomaterials.

[36]  Elisabeth Wong,et al.  MICROFLUIDIC PLATFORM FOR THE QUANTITATIVE ANALYSIS OF LEUKOCYTE MIGRATION SIGNATURES , 2014, Nature Communications.

[37]  A. Folch,et al.  Microfluidic transwell inserts for generation of tissue culture-friendly gradients in well plates. , 2014, Lab on a chip.

[38]  D. Beebe,et al.  Characterizing asthma from a drop of blood using neutrophil chemotaxis , 2014, Proceedings of the National Academy of Sciences.

[39]  Jim C. Cheng,et al.  Multi-temperature zone, droplet-based microreactor for increased temperature control in nanoparticle synthesis. , 2014, Small.

[40]  Mehmet Toner,et al.  Collective and Individual Migration following the Epithelial-Mesenchymal Transition , 2014, Nature materials.

[41]  Chien-Chung Peng,et al.  A polydimethylsiloxane-polycarbonate hybrid microfluidic device capable of generating perpendicular chemical and oxygen gradients for cell culture studies. , 2014, Lab on a chip.

[42]  Lidong Qin,et al.  Mesenchymal-mode migration assay and antimetastatic drug screening with high-throughput microfluidic channel networks. , 2014, Angewandte Chemie.

[43]  G. Dubini,et al.  Human 3D vascularized organotypic microfluidic assays to study breast cancer cell extravasation , 2014, Proceedings of the National Academy of Sciences.

[44]  Josep Puigmartí-Luis,et al.  Microfluidic platforms: a mainstream technology for the preparation of crystals. , 2014, Chemical Society reviews.

[45]  D. Beebe,et al.  Simple microfluidic device for studying chemotaxis in response to dual gradients , 2015, Biomedical microdevices.

[46]  A. Aranyosi,et al.  Microfluidic mazes to characterize T-cell exploration patterns following activation in vitro. , 2015, Integrative biology : quantitative biosciences from nano to macro.

[47]  T. Konry,et al.  Phenotypic drug profiling in droplet microfluidics for better targeting of drug-resistant tumors. , 2015, Lab on a chip.

[48]  A. Olivares,et al.  Generation of stable orthogonal gradients of chemical concentration and substrate stiffness in a microfluidic device. , 2015, Lab on a chip.

[49]  Junbo Wang,et al.  A Tubing-Free Microfluidic Wound Healing Assay Enabling the Quantification of Vascular Smooth Muscle Cell Migration , 2015, Scientific Reports.

[50]  Evan Z. Macosko,et al.  Highly Parallel Genome-wide Expression Profiling of Individual Cells Using Nanoliter Droplets , 2015, Cell.

[51]  T E de Groot,et al.  Surface-tension driven open microfluidic platform for hanging droplet culture. , 2016, Lab on a chip.

[52]  C. Luo,et al.  Gel integration for microfluidic applications. , 2016, Lab on a chip.

[53]  Brian F. Bender,et al.  Digital microfluidics for spheroid-based invasion assays. , 2016, Lab on a chip.

[54]  Nitin Agrawal,et al.  Development of a Single-Cell Migration and Extravasation Platform through Selective Surface Modification. , 2016, Analytical chemistry.

[55]  Noo Li Jeon,et al.  Three-dimensional biomimetic model to reconstitute sprouting lymphangiogenesis in vitro. , 2016, Biomaterials.

[56]  Mehmet Toner,et al.  Clusters of circulating tumor cells traverse capillary-sized vessels , 2016, Proceedings of the National Academy of Sciences.

[57]  Daniel Irimia,et al.  Big insights from small volumes: deciphering complex leukocyte behaviors using microfluidics , 2016, Journal of leukocyte biology.