Microfluidics for 3D Cell and Tissue Cultures: Microfabricative and Ethical Aspects Updates

The necessity to improve in vitro cell screening assays is becoming ever more important. Pharmaceutical companies, research laboratories and hospitals require technologies that help to speed up conventional screening and therapeutic procedures to produce more data in a short time in a realistic and reliable manner. The design of new solutions for test biomaterials and active molecules is one of the urgent problems of preclinical screening and the limited correlation between in vitro and in vivo data remains one of the major issues. The establishment of the most suitable in vitro model provides reduction in times, costs and, last but not least, in the number of animal experiments as recommended by the 3Rs (replace, reduce, refine) ethical guiding principles for testing involving animals. Although two-dimensional (2D) traditional cell screening assays are generally cheap and practical to manage, they have strong limitations, as cells, within the transition from the three-dimensional (3D) in vivo to the 2D in vitro growth conditions, do not properly mimic the real morphologies and physiology of their native tissues. In the study of human pathologies, especially, animal experiments provide data closer to what happens in the target organ or apparatus, but they imply slow and costly procedures and they generally do not fully accomplish the 3Rs recommendations, i.e., the amount of laboratory animals and the stress that they undergo must be minimized. Microfluidic devices seem to offer different advantages in relation to the mentioned issues. This review aims to describe the critical issues connected with the conventional cells culture and screening procedures, showing what happens in the in vivo physiological micro and nano environment also from a physical point of view. During the discussion, some microfluidic tools and their components are described to explain how these devices can circumvent the actual limitations described in the introduction.

[1]  Nam-Trung Nguyen,et al.  Microfluidic gut-on-a-chip with three-dimensional villi structure , 2017, Biomedical microdevices.

[2]  A. Waqas,et al.  Technological Advances of 3D Scaffold-Based Stem Cell/Exosome Therapy in Tissues and Organs , 2021, Frontiers in Cell and Developmental Biology.

[3]  Chengyan Zhang,et al.  3D Culture System for Liver Tissue Mimicking Hepatic Plates for Improvement of Human Hepatocyte (C3A) Function and Polarity , 2020, BioMed research international.

[4]  G. Zuccotti,et al.  Human endothelial cells in high glucose: New clues from culture in 3D microfluidic chips , 2022, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[5]  G. Eknoyan,et al.  Endothelial characteristics of glomerular capillaries in normal, mercuric chloride-induced, and gentamicin-induced acute renal failure in the rat. , 1983, The Journal of clinical investigation.

[6]  D. L. Taylor,et al.  A glass-based, continuously zonated and vascularized human liver acinus microphysiological system (vLAMPS) designed for experimental modeling of diseases and ADME/TOX. , 2018, Lab on a chip.

[7]  D. A. Severinov,et al.  Ethical and legal aspects of in vivo experimental biomedical research of the conduct. Part II , 2019, I.P. Pavlov Russian Medical Biological Herald.

[8]  G. Perozziello,et al.  A Disposable Passive Microfluidic Device for Cell Culturing , 2020, Biosensors.

[9]  M. Vaher,et al.  Miniaturization of sampling for chemical reaction monitoring by capillary electrophoresis. , 2005, Journal of chromatography. A.

[10]  Albert Gough,et al.  A human liver microphysiology platform for investigating physiology, drug safety, and disease models , 2016, Experimental biology and medicine.

[11]  V. Petrikaitė,et al.  3D Cell Culture Models as Recapitulators of the Tumor Microenvironment for the Screening of Anti-Cancer Drugs , 2021, Cancers.

[12]  Krist V. Gernaey,et al.  Lab on a chip automates in vitro cell culturing , 2012 .

[13]  Gang Bao,et al.  Tumour-on-a-chip: microfluidic models of tumour morphology, growth and microenvironment , 2017, Journal of The Royal Society Interface.

[14]  J. Kang,et al.  Collagen-based brain microvasculature model in vitro using three-dimensional printed template. , 2015, Biomicrofluidics.

[15]  Ehsan Samiei,et al.  A review of digital microfluidics as portable platforms for lab-on a-chip applications. , 2016, Lab on a chip.

[16]  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.

[17]  G. Perozziello,et al.  Development of 3D PVA scaffolds for cardiac tissue engineering and cell screening applications , 2019, RSC advances.

[18]  Teck Chuan Lim,et al.  A microfluidic 3D hepatocyte chip for drug toxicity testing. , 2009, Lab on a chip.

[19]  M. Nikkhah,et al.  Developing 3D Organized Human Cardiac Tissue within a Microfluidic Platform. , 2021, Journal of visualized experiments : JoVE.

[20]  Benjamin S. Freedman,et al.  3D cell culture models: Drug pharmacokinetics, safety assessment, and regulatory consideration , 2021, Clinical and translational science.

[21]  R. Voronov,et al.  A Minireview of Microfluidic Scaffold Materials in Tissue Engineering , 2022, Frontiers in Molecular Biosciences.

[22]  Roger D. Kamm,et al.  A 3D neurovascular microfluidic model consisting of neurons, astrocytes and cerebral endothelial cells as a blood-brain barrier. , 2017, Lab on a chip.

[23]  Martin Engel,et al.  Generation and Analysis of 3D Cell Culture Models for Drug Discovery. , 2021, European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences.

[24]  Ying Bai,et al.  Bioactive Decellularized Extracellular Matrix Hydrogel Microspheres Fabricated Using a Temperature-Controlling Microfluidic System. , 2022, ACS biomaterials science & engineering.

[25]  Francesco De Angelis,et al.  A microfluidic device integrating plasmonic nanodevices for Raman spectroscopy analysis on trapped single living cells , 2013 .

[26]  G. Reilly,et al.  Design and Evaluation of an Osteogenesis-on-a-Chip Microfluidic Device Incorporating 3D Cell Culture , 2020, Frontiers in Bioengineering and Biotechnology.

[27]  C. Bouten,et al.  A biomimetic microfluidic model to study signalling between endothelial and vascular smooth muscle cells under hemodynamic conditions , 2018, Lab on a chip.

[28]  Rosanna La Rocca,et al.  Microfluidic biofunctionalisation protocols to form multi‐valent interactions for cell rolling and phenotype modification investigations , 2013, Electrophoresis.

[29]  C. Olgasi,et al.  iPSC-Derived Liver Organoids: A Journey from Drug Screening, to Disease Modeling, Arriving to Regenerative Medicine , 2020, International journal of molecular sciences.

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

[31]  Liwei Lin,et al.  3D printed microfluidic devices for circulating tumor cells (CTCs) isolation. , 2019, Biosensors & bioelectronics.

[32]  Hemant Sarin,et al.  Physiologic upper limits of pore size of different blood capillary types and another perspective on the dual pore theory of microvascular permeability , 2010, Journal of angiogenesis research.

[33]  Microfluidic Bioreactor Made of Cyclo-Olefin Polymer for Observing On-Chip Platelet Production , 2021, Micromachines.

[34]  W. Verdurmen,et al.  Oxygen control: the often overlooked but essential piece to create better in vitro systems. , 2022, Lab on a chip.

[35]  D. R. Myers,et al.  Vascularized Microfluidics and Their Untapped Potential for Discovery in Diseases of the Microvasculature , 2021, Annual review of biomedical engineering.

[36]  Jason P. Gleghorn,et al.  Microfluidic scaffolds for tissue engineering. , 2007, Nature materials.

[37]  Noo Li Jeon,et al.  High-Throughput Microfluidic 3D Cytotoxicity Assay for Cancer Immunotherapy (CACI-IMPACT Platform) , 2019, Front. Immunol..

[38]  Y. Tabata,et al.  A cancer invasion model combined with CAF aggregates incorporating GM containing a p53 inhibitor. , 2019, Tissue engineering. Part C, Methods.

[39]  Gad D Vatine,et al.  Microfluidic channel sensory system for electro-addressing cell location, determining confluency, and quantifying a general number of cells , 2022, Scientific Reports.

[40]  Gerardo Perozziello,et al.  Ca2+ Mediates the Adhesion of Breast Cancer Cells in Self-Assembled Multifunctional Microfluidic Chip Prepared with Carbohydrate Beads , 2010 .

[41]  Jeong‐Yeol Yoon,et al.  Microscopic Imaging Methods for Organ-on-a-Chip Platforms , 2022, Micromachines.

[42]  A. I. Gómez-Varela,et al.  Microfluidic devices manufacturing with a stereolithographic printer for biological applications. , 2021, Materials science & engineering. C, Materials for biological applications.

[43]  R. De Caro,et al.  Tissue-Engineered Grafts from Human Decellularized Extracellular Matrices: A Systematic Review and Future Perspectives , 2018, International journal of molecular sciences.

[44]  K. Rakušan,et al.  Capillary length, tortuosity, and spacing in rat myocardium during cardiac cycle. , 1992, The American journal of physiology.

[45]  N. Jeon,et al.  Perfusable micro-vascularized 3D tissue array for high-throughput vascular phenotypic screening , 2022, Nano Convergence.

[46]  Chapter 7 – Compartmental Modeling , 2012 .

[47]  Weijia Wen,et al.  Organ-on-a-chip: recent breakthroughs and future prospects , 2020, BioMedical Engineering OnLine.

[48]  Fabio Benfenati,et al.  Nanostructured superhydrophobic substrates trigger the development of 3D neuronal networks. , 2013, Small.

[49]  Andreas Manz,et al.  Galectin-3 coats the membrane of breast cells and makes a signature of tumours. , 2014, Molecular bioSystems.

[50]  B. Alberts,et al.  Blood Vessels and Endothelial Cells , 2002 .

[51]  R. Fleck,et al.  3D microfluidic liver cultures as a physiological preclinical tool for hepatitis B virus infection , 2018, Nature Communications.

[52]  Herman Goossens,et al.  3D culture of murine neural stem cells on decellularized mouse brain sections. , 2015, Biomaterials.

[53]  Dan Gao,et al.  Recent Development of Drug Delivery Systems through Microfluidics: From Synthesis to Evaluation , 2022, Pharmaceutics.

[54]  P. Ertl,et al.  A Microfluidic Multisize Spheroid Array for Multiparametric Screening of Anticancer Drugs and Blood–Brain Barrier Transport Properties , 2021, Advanced science.

[55]  Lukas Mathur,et al.  Microfluidics as an Enabling Technology for Personalized Cancer Therapy. , 2019, Small.

[56]  V. Mollaki Ethical Challenges in Organoid Use , 2021, Biotech (Basel (Switzerland)).

[57]  K. Lamperska,et al.  2D and 3D cell cultures – a comparison of different types of cancer cell cultures , 2016, Archives of medical science : AMS.

[58]  Evaluation and optimization of PolyJet 3D-printed materials for cell culture studies , 2022, Analytical and Bioanalytical Chemistry.

[59]  Noo Li Jeon,et al.  Recreating the perivascular niche ex vivo using a microfluidic approach , 2010, Biotechnology and bioengineering.

[60]  Z. Nie,et al.  Microfluidic 3D cell culture: potential application for tissue-based bioassays. , 2012, Bioanalysis.

[61]  N. Jeon,et al.  3D Microfluidic Bone Tumor Microenvironment Comprised of Hydroxyapatite/Fibrin Composite , 2019, Front. Bioeng. Biotechnol..

[62]  Fabio Benfenati,et al.  Delivery of Brain-Derived Neurotrophic Factor by 3D Biocompatible Polymeric Scaffolds for Neural Tissue Engineering and Neuronal Regeneration , 2018, Molecular Neurobiology.

[63]  A. Papadimitropoulos,et al.  Maintenance of Primary Human Colorectal Cancer Microenvironment Using a Perfusion Bioreactor‐Based 3D Culture System , 2019, Advanced biosystems.

[64]  H. Kim,et al.  3D in vitro morphogenesis of human intestinal epithelium in a gut-on-a-chip or a hybrid chip with a cell culture insert , 2022, Nature Protocols.

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

[66]  Xiaofeng Jia,et al.  Three-dimensional (3D) printed scaffold and material selection for bone repair. , 2019, Acta biomaterialia.

[67]  Ishan Pandey,et al.  Transcending toward Advanced 3D-Cell Culture Modalities: A Review about an Emerging Paradigm in Translational Oncology , 2021, Cells.

[68]  Ronan M. T. Fleming,et al.  Automated microuidic cell culture of stem cell derived dopaminergic neurons in Parkinson’s disease , 2017, bioRxiv.

[69]  Anthony D. Saleh,et al.  A 3D Microfluidic Liver Model for High Throughput Compound Toxicity Screening in the OrganoPlate®. , 2020, Toxicology.

[70]  V. Baumans,et al.  Use of animals in experimental research: an ethical dilemma? , 2004, Gene therapy.

[71]  Nipha Chaicharoenaudomrung,et al.  Three-dimensional cell culture systems as an in vitro platform for cancer and stem cell modeling , 2019, World journal of stem cells.

[72]  J. Choi,et al.  Wnt5a-mediating neurogenesis of human adipose tissue-derived stem cells in a 3D microfluidic cell culture system. , 2011, Biomaterials.

[73]  Hwa Liang Leo,et al.  A 3D printed microfluidic perfusion device for multicellular spheroid cultures , 2017, Biofabrication.

[74]  Philippe Bédard,et al.  Innovative Human Three-Dimensional Tissue-Engineered Models as an Alternative to Animal Testing , 2020, Bioengineering.

[75]  A. Woolley,et al.  Advances in microfluidic materials, functions, integration, and applications. , 2013, Chemical reviews.

[76]  Yong Teng,et al.  Is It Time to Start Transitioning From 2D to 3D Cell Culture? , 2020, Frontiers in Molecular Biosciences.

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

[78]  Sang-Hoon Lee,et al.  A 3D alcoholic liver disease model on a chip. , 2016, Integrative biology : quantitative biosciences from nano to macro.

[79]  Alicia C B Allen,et al.  Multilayer microfluidic PEGDA hydrogels. , 2010, Biomaterials.

[80]  D. H. Kim,et al.  In Vitro Models Mimicking Immune Response in the Skin , 2021, Yonsei medical journal.

[81]  M. Ganjali,et al.  Human Organs-on-Chips: A Review of the State-of-the-Art, Current Prospects, and Future Challenges. , 2021, Advanced biology.

[82]  Joseph Wang,et al.  On‐chip enzymatic assays , 2002, Electrophoresis.

[83]  Noo Li Jeon,et al.  "Open-top" microfluidic device for in vitro three-dimensional capillary beds. , 2017, Lab on a chip.

[84]  Yi Zhang,et al.  3D-printed Bioreactors for In Vitro Modeling and Analysis , 2020, International journal of bioprinting.

[85]  C. Estill,et al.  Matrix-free three-dimensional culture of bovine secondary follicles to antral stage: Impact of media formulation and epidermal growth factor (EGF). , 2022, Theriogenology.

[86]  Sigrid A. Langhans Three-Dimensional in Vitro Cell Culture Models in Drug Discovery and Drug Repositioning , 2018, Front. Pharmacol..

[87]  Hidetoshi Kotera,et al.  Integrating perfusable vascular networks with a three-dimensional tissue in a microfluidic device. , 2017, Integrative biology : quantitative biosciences from nano to macro.

[88]  M. Sur,et al.  A low-cost 3D printed microfluidic bioreactor and imaging chamber for live-organoid imaging. , 2021, Biomicrofluidics.

[89]  Courtney M. Sakolish,et al.  Analysis of Reproducibility and Robustness of a Human Microfluidic Four-Cell Liver Acinus MicroPhysiology System (LAMPS). , 2020, Toxicology.

[90]  G. Perozziello,et al.  A Passive Microfluidic Device for Chemotaxis Studies , 2019, Micromachines.

[91]  Y. Tabata,et al.  Three-Dimensional Culture System of Cancer Cells Combined with Biomaterials for Drug Screening , 2020, Cancers.

[92]  N. Manaresi,et al.  A microvalve for hybrid microfluidic systems , 2010, DTIP 2010.

[93]  Maddaly Ravi,et al.  3D Cell Culture Systems: Advantages and Applications , 2015, Journal of cellular physiology.

[94]  P. Ertl,et al.  Recent Advances in Additive Manufacturing and 3D Bioprinting for Organs-On-A-Chip and Microphysiological Systems , 2022, Frontiers in Bioengineering and Biotechnology.

[95]  X. Mu,et al.  Design and fabrication of a liver-on-a-chip platform for convenient, highly efficient, and safe in situ perfusion culture of 3D hepatic spheroids. , 2018, Lab on a chip.

[96]  G. Whitesides,et al.  A paper-based invasion assay: assessing chemotaxis of cancer cells in gradients of oxygen. , 2015, Biomaterials.

[97]  Jing Liu,et al.  Engineered 3D tumour model for study of glioblastoma aggressiveness and drug evaluation on a detachably assembled microfluidic device , 2018, Biomedical Microdevices.

[98]  M. Ravi,et al.  Devices and techniques used to obtain and analyze three‐dimensional cell cultures , 2021, Biotechnology progress.

[99]  Marco Rasponi,et al.  Bioprinting 3D microfibrous scaffolds for engineering endothelialized myocardium and heart-on-a-chip. , 2016, Biomaterials.

[100]  E. Gottwald,et al.  Advanced 3D Cell Culture Techniques in Micro-Bioreactors, Part I: A Systematic Analysis of the Literature Published between 2000 and 2020 , 2020, Processes.

[101]  U. Losert,et al.  Introducing the concept of the 3Rs into tissue engineering research. , 2006, ALTEX.

[102]  Detlef Snakenborg,et al.  A fast and reliable way to establish fluidic connections to planar microchips , 2006 .

[103]  R. Fair,et al.  An integrated digital microfluidic lab-on-a-chip for clinical diagnostics on human physiological fluids. , 2004, Lab on a chip.

[104]  G. Perozziello,et al.  Fabrication and Applications of Micro/Nanostructured Devices for Tissue Engineering , 2016, Nano-Micro Letters.