Microfluidic Technology for the Generation of Cell Spheroids and Their Applications

A three-dimensional (3D) tissue model has significant advantages over the conventional two-dimensional (2D) model. A 3D model mimics the relevant in-vivo physiological conditions, allowing a cell culture to serve as an effective tool for drug discovery, tissue engineering, and the investigation of disease pathology. The present reviews highlight the recent advances and the development of microfluidics based methods for the generation of cell spheroids. The paper emphasizes on the application of microfluidic technology for tissue engineering including the formation of multicellular spheroids (MCS). Further, the paper discusses the recent technical advances in the integration of microfluidic devices for MCS-based high-throughput drug screening. The review compares the various microfluidic techniques and finally provides a perspective for the future opportunities in this research area.

[1]  Frantisek Foret,et al.  Droplet microfluidics in (bio)chemical analysis. , 2015, The Analyst.

[2]  Youmin Hou,et al.  In Vitro Epithelial Organoid Generation Induced by Substrate Nanotopography , 2015, Scientific Reports.

[3]  Weixin Zhao,et al.  Microencapsulation of porcine thyroid cell organoids within a polymer microcapsule construct , 2017, Experimental biology and medicine.

[4]  A. Kouzani,et al.  Microfluidic devices for cell cultivation and proliferation. , 2013, Biomicrofluidics.

[5]  Samin Akbari,et al.  Microfluidic encapsulation of cells in alginate particles via an improved internal gelation approach , 2013, Microfluidics and Nanofluidics.

[6]  Kenji Suzuki,et al.  Electrowetting-Based Actuation of Liquid Droplets for Micro Transportation Systems , 2010 .

[7]  C. Archer,et al.  Pellet culture model for human primary osteoblasts. , 2010, European cells & materials.

[8]  P. Vos,et al.  Cell encapsulation: Promise and progress , 2003, Nature Medicine.

[9]  Wei Shen,et al.  A novel technique for the formation of embryoid bodies inside liquid marbles , 2013 .

[10]  J. Baret.  Surfactants in droplet-based microfluidics. , 2012, Lab on a chip.

[11]  A. Wheeler,et al.  Digital microfluidics for cell-based assays. , 2008, Lab on a chip.

[12]  G. Rice,et al.  α2β1 integrin affects metastatic potential of ovarian carcinoma spheroids by supporting disaggregation and proteolysis , 2007, Journal of Carcinogenesis.

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

[14]  Christoph A. Merten,et al.  Droplet-based microfluidic platforms for the encapsulation and screening of Mammalian cells and multicellular organisms. , 2008, Chemistry & biology.

[15]  Michael R. King,et al.  Continuously perfused microbubble array for 3D tumor spheroid model. , 2011, Biomicrofluidics.

[16]  S. Bryant,et al.  Cell encapsulation in biodegradable hydrogels for tissue engineering applications. , 2008, Tissue engineering. Part B, Reviews.

[17]  Subra Suresh,et al.  Three-dimensional manipulation of single cells using surface acoustic waves , 2016, Proceedings of the National Academy of Sciences.

[18]  B. Harley,et al.  The use of covalently immobilized stem cell factor to selectively affect hematopoietic stem cell activity within a gelatin hydrogel. , 2015, Biomaterials.

[19]  Tao Wang,et al.  Surface Acoustic Waves (SAW)-Based Biosensing for Quantification of Cell Growth in 2D and 3D Cultures , 2015, Sensors.

[20]  P. Lee,et al.  Microfluidic array for three-dimensional perfusion culture of human mammary epithelial cells , 2011, Biomedical microdevices.

[21]  Juergen Friedrich,et al.  Experimental anti-tumor therapy in 3-D: Spheroids – old hat or new challenge? , 2007, International journal of radiation biology.

[22]  J. Palefsky,et al.  Role of E-cadherin in the induction of apoptosis of HPV16-positive CaSki cervical cancer cells during multicellular tumor spheroid formation , 2007, Apoptosis.

[23]  D. Weitz,et al.  Monodisperse Double Emulsions Generated from a Microcapillary Device , 2005, Science.

[24]  Yongsheng Su,et al.  Surface Tension Guided Hanging-Drop: Producing Controllable 3D Spheroid of High-Passaged Human Dermal Papilla Cells and Forming Inductive Microtissues for Hair-Follicle Regeneration. , 2016, ACS applied materials & interfaces.

[25]  W. Birchmeier,et al.  E-cadherin-mediated cell-cell adhesion prevents invasiveness of human carcinoma cells , 1991, The Journal of cell biology.

[26]  R. Garrell,et al.  Digital Microfluidics for Automated Hanging Drop Cell Spheroid Culture , 2015, Journal of laboratory automation.

[27]  Yuan Wen,et al.  The future of microfluidic assays in drug development , 2008, Expert opinion on drug discovery.

[28]  C. Cho,et al.  Role of E-cadherin Molecules in Spheroid Formation of Hepatocytes Adhered on Galactose-Carrying Polymer as an Artificial Asialoglycoprotein Model , 2005, Biotechnology Letters.

[29]  Juan Xu,et al.  Simple and reusable off-the-shelf microfluidic devices for the versatile generation of droplets. , 2016, Lab on a chip.

[30]  Shuichi Takayama,et al.  High-throughput 3D spheroid culture and drug testing using a 384 hanging drop array. , 2011, The Analyst.

[31]  S. Cho,et al.  Creating, transporting, cutting, and merging liquid droplets by electrowetting-based actuation for digital microfluidic circuits , 2003 .

[32]  Utkan Demirci,et al.  Biotunable Acoustic Node Assembly of Organoids , 2015, Advanced healthcare materials.

[33]  D. Weitz,et al.  Microfluidic fabrication of monodisperse biocompatible and biodegradable polymersomes with controlled permeability. , 2008, Journal of the American Chemical Society.

[34]  William L. Haisler,et al.  Three-dimensional cell culturing by magnetic levitation , 2013, Nature Protocols.

[35]  K. S. Narayan,et al.  Three-dimensional growth patterns of various human tumor cell lines in simulated microgravity of a NASA bioreactor , 1997, In Vitro Cellular & Developmental Biology - Animal.

[36]  J. Morgan,et al.  Quantifying the kinetics and morphological changes of the fusion of spheroid building blocks , 2016, Biofabrication.

[37]  Shoji Takeuchi,et al.  Encapsulating bacteria in agarose microparticles using microfluidics for high-throughput cell analysis and isolation. , 2011, ACS chemical biology.

[38]  Chwee Teck Lim,et al.  Microfluidic cell trap array for controlled positioning of single cells on adhesive micropatterns. , 2013, Lab on a chip.

[39]  Mandy B. Esch,et al.  How multi-organ microdevices can help foster drug development. , 2014, Advanced drug delivery reviews.

[40]  Sang Joon Lee,et al.  Three-dimensional digital microfluidic manipulation of droplets in oil medium , 2015, Scientific Reports.

[41]  Jinyi Wang,et al.  Mixed hydrogel bead-based tumor spheroid formation and anticancer drug testing. , 2014, The Analyst.

[42]  Eugenia Kumacheva,et al.  Chitosan/agarose hydrogels: cooperative properties and microfluidic preparation. , 2014, Carbohydrate polymers.

[43]  Jiehong Liao,et al.  Bioactive polymer/extracellular matrix scaffolds fabricated with a flow perfusion bioreactor for cartilage tissue engineering. , 2010, Biomaterials.

[44]  Andreas Hierlemann,et al.  Reconfigurable microfluidic hanging drop network for multi-tissue interaction and analysis , 2014, Nature Communications.

[45]  Xudong Shi,et al.  Biodegradable polymeric microcarriers with controllable porous structure for tissue engineering. , 2009, Macromolecular bioscience.

[46]  Mingwen Ma,et al.  Functional three‐dimensional HepG2 aggregate cultures generated from an ultrasound trap: Comparison with HepG2 spheroids , 2007, Journal of cellular biochemistry.

[47]  Thomas Hankemeier,et al.  Microfluidic 3D cell culture: from tools to tissue models. , 2015, Current opinion in biotechnology.

[48]  James Briscoe,et al.  The interpretation of morphogen gradients , 2006, Development.

[49]  Scott D. Soelberg,et al.  Microfluidic one-step synthesis of alginate microspheres immobilized with antibodies , 2013, Journal of The Royal Society Interface.

[50]  B. Godin,et al.  3D In Vitro Model for Breast Cancer Research Using Magnetic Levitation and Bioprinting Method. , 2016, Methods in molecular biology.

[51]  K. Cheung,et al.  Droplet-based microfluidic system for multicellular tumor spheroid formation and anticancer drug testing. , 2010, Lab on a chip.

[52]  R. Fair,et al.  Electrowetting-based actuation of liquid droplets for microfluidic applications , 2000 .

[53]  Ying Zhang,et al.  Efficient One-Step Production of Microencapsulated Hepatocyte Spheroids with Enhanced Functions. , 2016, Small.

[54]  S. Quake,et al.  Microfluidic Large-Scale Integration , 2002, Science.

[55]  D. Weitz,et al.  Dripping, Jetting, Drops, and Wetting: The Magic of Microfluidics , 2007 .

[56]  S. Cerdán,et al.  Microscopic images of intraspheroidal pH by 1H magnetic resonance chemical shift imaging of pH sensitive indicators , 2005, Magnetic Resonance Materials in Physics, Biology and Medicine.

[57]  S. Sugiura,et al.  Detachably assembled microfluidic device for perfusion culture and post-culture analysis of a spheroid array. , 2014, Biotechnology journal.

[58]  Ali Khademhosseini,et al.  Microfluidic patterning for fabrication of contractile cardiac organoids , 2007, Biomedical microdevices.

[59]  Rui Zhang,et al.  Rapid formation of size-controllable multicellular spheroids via 3D acoustic tweezers. , 2016, Lab on a chip.

[60]  N. Nguyen,et al.  A lab-on-a-chip device for investigating the fusion process of olfactory ensheathing cell spheroids. , 2016, Lab on a chip.

[61]  Hanry Yu,et al.  A practical guide to microfluidic perfusion culture of adherent mammalian cells. , 2007, Lab on a chip.

[62]  Biana Godin,et al.  Three-Dimensional In Vitro Co-Culture Model of Breast Tumor using Magnetic Levitation , 2014, Scientific Reports.

[63]  G. Darlington,et al.  Induction of three-dimensional assembly of human liver cells by simulated microgravity , 1999, In Vitro Cellular & Developmental Biology - Animal.

[64]  Hans Clevers,et al.  Organoids: Modeling Development and the Stem Cell Niche in a Dish. , 2016, Developmental cell.

[65]  A. Ivascu,et al.  Rapid Generation of Single-Tumor Spheroids for High-Throughput Cell Function and Toxicity Analysis , 2006, Journal of biomolecular screening.

[66]  N. Hibino,et al.  Principles of the Kenzan Method for Robotic Cell Spheroid-Based 3 D Bioprinting , 2016 .

[67]  Dongyue Jiang,et al.  Light-driven 3D droplet manipulation on flexible optoelectrowetting devices fabricated by a simple spin-coating method. , 2016, Lab on a chip.

[68]  T. Aigner,et al.  Micromass co-culture of human articular chondrocytes and human bone marrow mesenchymal stem cells to investigate stable neocartilage tissue formation in vitro. , 2010, European cells & materials.

[69]  L. Álvarez-Vallina,et al.  Microencapsulation of therapeutic bispecific antibodies producing cells: immunotherapeutic organoids for cancer management , 2015, Journal of drug targeting.

[70]  Shuichi Takayama,et al.  Transfer, Imaging, and Analysis Plate for Facile Handling of 384 Hanging Drop 3D Tissue Spheroids , 2014, Journal of laboratory automation.

[71]  Ivan Martin,et al.  Use of multicellular tumor spheroids to dissect endothelial cell–tumor cell interactions: A role for T‐cadherin in tumor angiogenesis , 2007, FEBS letters.

[72]  Robert Langer,et al.  Microfluidic platform for controlled synthesis of polymeric nanoparticles. , 2008, Nano letters.

[73]  Kelly M. Schultz,et al.  High-throughput rheology in a microfluidic device. , 2011, Lab on a chip.

[74]  R. Shatford,et al.  Extended liver-specific functions of porcine hepatocyte spheroids entrapped in collagen gel , 1995, In Vitro Cellular & Developmental Biology - Animal.

[75]  V. Torchilin,et al.  Generation and functional assessment of 3D multicellular spheroids in droplet based microfluidics platform. , 2016, Lab on a chip.

[76]  W. Mueller‐Klieser.  Microelectrode measurement of oxygen tension distributions in multicellular spheroids cultured in spinner flasks. , 1984, Recent results in cancer research. Fortschritte der Krebsforschung. Progres dans les recherches sur le cancer.

[77]  Faisal Mohd-Yasin,et al.  Digital microfluidics with a magnetically actuated floating liquid marble. , 2016, Lab on a chip.

[78]  Nam-Trung Nguyen,et al.  Design, fabrication and characterization of drug delivery systems based on lab-on-a-chip technology. , 2013, Advanced drug delivery reviews.

[79]  R. Kamm,et al.  Microfluidic models of vascular functions. , 2012, Annual review of biomedical engineering.

[80]  Y. Rhee,et al.  Generation of monodisperse alginate microbeads and in situ encapsulation of cell in microfluidic device , 2007, Biomedical microdevices.

[81]  J.V.L. da Silva,et al.  Scalable robotic biofabrication of tissue spheroids , 2011, Biofabrication.

[82]  Dietmar W. Hutmacher,et al.  Bioengineered 3D platform to explore cell-ECM interactions and drug resistance of epithelial ovarian cancer cells. , 2010, Biomaterials.

[83]  Renu John,et al.  Optofluidic bioimaging platform for quantitative phase imaging of lab on a chip devices using digital holographic microscopy. , 2016, Applied optics.

[84]  Emanuele Marconi,et al.  A microfluidic platform for chemoresistive testing of multicellular pleural cancer spheroids. , 2014, Lab on a chip.

[85]  Jeong‐Yeol Yoon,et al.  Preventing Biomolecular Adsorption in Electrowetting-Based Biofluidic Chips. , 2003, Analytical chemistry.

[86]  Kenneth M. Yamada,et al.  Taking Cell-Matrix Adhesions to the Third Dimension , 2001, Science.

[87]  N. Marceau,et al.  Spheroidal aggregate culture of rat liver cells: histotypic reorganization, biomatrix deposition, and maintenance of functional activities , 1985, The Journal of cell biology.

[88]  S. Anna,et al.  Coalescence and splitting of confined droplets at microfluidic junctions. , 2009, Lab on a chip.

[89]  Douglas W. DeSimone,et al.  Integrin-ECM Interactions Regulate Cadherin-Dependent Cell Adhesion and Are Required for Convergent Extension in Xenopus , 2003, Current Biology.

[90]  Dongqing Li,et al.  Three-dimensional modeling of transport of nutrients for multicellular tumor spheroid culture in a microchannel , 2007, Biomedical microdevices.

[91]  Junji Fukuda,et al.  An oxygen-permeable spheroid culture system for the prevention of central hypoxia and necrosis of spheroids. , 2012, Biomaterials.

[92]  V. Cristini,et al.  Scalings for Droplet Sizes in Shear-Driven Breakup: Non-Microfluidic Ways to Monodisperse Emulsions , 2006 .

[93]  Aaron R Wheeler,et al.  Hydrogel discs for digital microfluidics. , 2012, Biomicrofluidics.

[94]  Mattias Goksör,et al.  Optical tweezers applied to a microfluidic system. , 2004, Lab on a chip.

[95]  W. Mueller‐Klieser,et al.  Method for the determination of oxygen consumption rates and diffusion coefficients in multicellular spheroids. , 1984, Biophysical journal.

[96]  A. Khademhosseini,et al.  Preparation of arrays of cell spheroids and spheroid-monolayer cocultures within a microfluidic device. , 2010, Journal of bioscience and bioengineering.

[97]  A MOSCONA,et al.  The dissociation and aggregation of cells from organ rudiments of the early chick embryo. , 1952, Journal of anatomy.

[98]  Shuichi Takayama,et al.  Micro-ring structures stabilize microdroplets to enable long term spheroid culture in 384 hanging drop array plates , 2011, Biomedical Microdevices.

[99]  Hiang Kwee Lee,et al.  Spinning Liquid Marble and Its Dual Applications as Microcentrifuge and Miniature Localized Viscometer. , 2016, ACS applied materials & interfaces.

[100]  Mina Okochi,et al.  Magnetic force-based cell patterning for evaluation of the effect of stromal fibroblasts on invasive capacity in 3D cultures. , 2013, Biosensors & bioelectronics.

[101]  Mark W. Tibbitt,et al.  Hydrogels as extracellular matrix mimics for 3D cell culture. , 2009, Biotechnology and bioengineering.

[102]  F. Sonntag,et al.  A four-organ-chip for interconnected long-term co-culture of human intestine, liver, skin and kidney equivalents. , 2015, Lab on a chip.

[103]  E. Kumacheva,et al.  Microfluidic encapsulation of cells in polymer microgels. , 2012, Small.

[104]  Sai Siva Gorthi,et al.  Automated cell viability assessment using a microfluidics based portable imaging flow analyzer. , 2015, Biomicrofluidics.

[105]  Shuichi Takayama,et al.  Microfluidic system for formation of PC-3 prostate cancer co-culture spheroids. , 2009, Biomaterials.

[106]  R. Raphael,et al.  Assembly of a three-dimensional multitype bronchiole coculture model using magnetic levitation. , 2013, Tissue engineering. Part C, Methods.

[107]  Ning Zhang,et al.  Magnetic-directed patterning of cell spheroids. , 2014, Journal of biomedical materials research. Part A.

[108]  Q. Lin,et al.  A Mechanically Tunable Microfluidic Cell-Trapping Device. , 2014, Sensors and actuators. A, Physical.

[109]  Luke P. Lee,et al.  Microfluidic self-assembly of tumor spheroids for anticancer drug discovery , 2008, Biomedical microdevices.

[110]  M. Iwamoto,et al.  Terminal differentiation and calcification in rabbit chondrocyte cultures grown in centrifuge tubes: regulation by transforming growth factor beta and serum factors. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[111]  James A Bankson,et al.  Three-dimensional tissue culture based on magnetic cell levitation. , 2010, Nature nanotechnology.

[112]  Bahram Javidi,et al.  Optofluidic system for three-dimensional sensing and identification of micro-organisms with digital holographic microscopy. , 2010, Optics letters.

[113]  Oleg Gendelman,et al.  A floating self-propelling liquid marble containing aqueous ethanol solutions , 2015 .

[114]  Gerard H Markx,et al.  Tissue engineering with electric fields: Immobilization of mammalian cells in multilayer aggregates using dielectrophoresis , 2007, Biotechnology and bioengineering.

[115]  Alar Ainla,et al.  A microfluidic pipette for single-cell pharmacology. , 2010, Analytical chemistry.

[116]  I. Sandu,et al.  The influence of gravity on the distribution of the deposit formed onto a substrate by sessile, hanging, and sandwiched hanging drop evaporation. , 2011, Journal of colloid and interface science.

[117]  J. Vasiliev,et al.  Rearrangements of the Actin Cytoskeleton and E-Cadherin–Based Adherens Junctions Caused by Neoplasic Transformation Change Cell–Cell Interactions , 2009, PloS one.

[118]  D. Baigl,et al.  Photo-actuation of liquids for light-driven microfluidics: state of the art and perspectives. , 2012, Lab on a chip.

[119]  R. Mrowka,et al.  Generation of Multicellular Breast Cancer Tumor Spheroids: Comparison of Different Protocols , 2016, Journal of Mammary Gland Biology and Neoplasia.

[120]  Sung Yong Park,et al.  Light-Driven Droplet Manipulation Technologies for Lab-on-a-Chip Applications , 2011 .

[121]  Despina Bazou,et al.  Physical enviroment of 2-D animal cell aggregates formed in a short pathlength ultrasound standing wave trap. , 2005, Ultrasound in medicine & biology.

[122]  G Hamilton,et al.  Multicellular spheroids as an in vitro tumor model. , 1998, Cancer letters.

[123]  A. Bosserhoff,et al.  Development of a model system to analyze chondrogenic differentiation of mesenchymal stem cells. , 2013, International journal of clinical and experimental pathology.

[124]  M. Ramasubramanian,et al.  A three-dimensional microfluidic approach to scaling up microencapsulation of cells , 2012, Biomedical Microdevices.

[125]  Michele Zagnoni,et al.  Transitioning from multi-phase to single-phase microfluidics for long-term culture and treatment of multicellular spheroids. , 2016, Lab on a chip.

[126]  Ya-Tang Yang,et al.  A nanoliter microfluidic serial dilution bioreactor. , 2015, Biomicrofluidics.

[127]  Jimmy Ly,et al.  Automated Reagent-Dispensing System for Microfluidic Cell Biology Assays , 2013, Journal of laboratory automation.

[128]  Yuanjin Zhao,et al.  Microfluidic Generation of Porous Microcarriers for Three-Dimensional Cell Culture. , 2015, ACS applied materials & interfaces.

[129]  Hwan-You Chang,et al.  Dynamic analysis of hepatoma spheroid formation: roles of E-cadherin and β1-integrin , 2006, Cell and Tissue Research.

[130]  Minseok S. Kim,et al.  A microfluidic platform for 3-dimensional cell culture and cell-based assays , 2007, Biomedical microdevices.

[131]  S. Bhatia,et al.  Engineering a perfusable 3D human liver platform from iPS cells. , 2016, Lab on a chip.

[132]  Yuriko Renardy,et al.  The effects of confinement and inertia on the production of droplets , 2006 .

[133]  Yong Zhang,et al.  Sustained release of hydrophobic drugs by the microfluidic assembly of multistage microgel/poly (lactic-co-glycolic acid) nanoparticle composites. , 2015, Biomicrofluidics.

[134]  Nicolas Bremond,et al.  Cellular capsules as a tool for multicellular spheroid production and for investigating the mechanics of tumor progression in vitro , 2013, Proceedings of the National Academy of Sciences.

[135]  G. Devi,et al.  Three-dimensional culture systems in cancer research: Focus on tumor spheroid model. , 2016, Pharmacology & therapeutics.

[136]  S. Takayama,et al.  Opportunities and challenges for use of tumor spheroids as models to test drug delivery and efficacy. , 2012, Journal of controlled release : official journal of the Controlled Release Society.

[137]  J. Park,et al.  Continuous generation of hydrogel beads and encapsulation of biological materials using a microfluidic droplet-merging channel , 2008 .

[138]  Junbo Wang,et al.  Development of Droplet Microfluidics Enabling High-Throughput Single-Cell Analysis , 2016, Molecules.

[139]  D. Grier.  A revolution in optical manipulation , 2003, Nature.

[140]  Chien-Chung Peng,et al.  A microfluidic device for uniform-sized cell spheroids formation, culture, harvesting and flow cytometry analysis. , 2013, Biomicrofluidics.

[141]  Andrew J deMello,et al.  Droplet-based microfluidics for artificial cell generation: a brief review , 2016, Interface Focus.

[142]  R. Reis,et al.  Liquid Marbles for High‐Throughput Biological Screening of Anchorage‐Dependent Cells , 2015, Advanced healthcare materials.

[143]  L. G. de la Torre,et al.  Microfluidic tools toward industrial biotechnology , 2016, Biotechnology progress.

[144]  Piero Rinaldo,et al.  Rapid, large‐scale formation of porcine hepatocyte spheroids in a novel spheroid reservoir bioartificial liver , 2005, Liver transplantation : official publication of the American Association for the Study of Liver Diseases and the International Liver Transplantation Society.

[145]  Q. Fang,et al.  Analytical detection techniques for droplet microfluidics--a review. , 2013, Analytica chimica acta.

[146]  L. Yeo,et al.  Continuous Production of Janus and Composite Liquid Marbles with Tunable Coverage. , 2016, ACS applied materials & interfaces.

[147]  M.C. Wu,et al.  Droplet Manipulation With Light on Optoelectrowetting Device , 2008, Journal of Microelectromechanical Systems.

[148]  G. Keller,et al.  In vitro differentiation of embryonic stem cells. , 1995, Current opinion in cell biology.

[149]  Juergen Friedrich,et al.  Spheroid-based drug screen: considerations and practical approach , 2009, Nature Protocols.

[150]  Cheng-Hsien Liu,et al.  A microfluidic chip with a U-shaped microstructure array for multicellular spheroid formation, culturing and analysis , 2014, Biofabrication.

[151]  Mark K. Goldberg,et al.  Performance Characterization of a Reconfigurable Planar-Array Digital Microfluidic System , 2006, IEEE Transactions on Computer-Aided Design of Integrated Circuits and Systems.

[152]  Uwe Marx,et al.  The Multi-organ Chip - A Microfluidic Platform for Long-term Multi-tissue Coculture , 2015, Journal of visualized experiments : JoVE.

[153]  Ethan Tumarkin,et al.  Microfluidic generation of microgels from synthetic and natural polymers. , 2009, Chemical Society reviews.

[154]  I. Giménez,et al.  In vitro systems to study nephropharmacology: 2D versus 3D models. , 2016, European journal of pharmacology.

[155]  N. Nguyen,et al.  Generation of three-dimensional multiple spheroid model of olfactory ensheathing cells using floating liquid marbles , 2015, Scientific Reports.

[156]  M. Sefton,et al.  Hepatic organoids for microfluidic drug screening. , 2014, Lab on a chip.

[157]  M. Tamura,et al.  Click-crosslinkable and photodegradable gelatin hydrogels for cytocompatible optical cell manipulation in natural environment , 2015, Scientific Reports.

[158]  D. Ingber,et al.  Microfluidic organs-on-chips , 2014, Nature Biotechnology.

[159]  Uwe Marx,et al.  A multi-organ chip co-culture of neurospheres and liver equivalents for long-term substance testing. , 2015, Journal of biotechnology.

[160]  H. Lundqvist,et al.  Adjuvant 131I-anti-CEA-antibody radioimmunotherapy inhibits the development of experimental colonic carcinoma liver metastases. , 1998, Anticancer Research.