Integrated multi-organ microphysiological systems are an evolving tool for preclinical evaluation of the potential toxicity and efficacy of drug candidates. Such systems, also known as Body-on-a-Chip devices, have a great potential to increase the successful conversion of drug candidates entering clinical trials into approved drugs. Systems, to be attractive for commercial adoption, need to be inexpensive, easy to operate, and give reproducible results. Further, the ability to measure functional responses, such as electrical activity, force generation, and barrier integrity of organ surrogates, enhances the ability to monitor response to drugs. The ability to operate a system for significant periods of time (up to 28 d) will provide potential to estimate chronic as well as acute responses of the human body. Here we review progress towards a self-contained low-cost microphysiological system with functional measurements of physiological responses.
Impact statement
Multi-organ microphysiological systems are promising devices to improve the drug development process. The development of a pumpless system represents the ability to build multi-organ systems that are of low cost, high reliability, and self-contained. These features, coupled with the ability to measure electrical and mechanical response in addition to chemical or metabolic changes, provides an attractive system for incorporation into the drug development process. This will be the most complete review of the pumpless platform with recirculation yet written.
EschMBSmithASProtJMOleagaCHickmanJJShulerML. How multi-organ microdevices can help foster drug development. Adv Drug Deliv Rev2014; 69–70: 158–69.
3.
EastwoodDFindlayLPooleSBirdCWadhwaMMooreMBurnsCThorpeRStebbingsR. Monoclonal antibody TGN1412 trial failure explained by species differences in CD28 expression on CD4+ effector memory T-cells. Br J Pharmacol2010; 161: 512–26.
4.
ShuaibALeesKRLydenPGrottaJDavalosADavisSMDienerHCAshwoodTWasiewskiWWEmeribeUInvestigatorsSIT. NXY-059 for the treatment of acute ischemic stroke. N Engl J Med2007; 357: 562–71.
5.
EschMBKingTLShulerML. The role of body-on-a-chip devices in drug and toxicity studies. Annu Rev Biomed Eng2011; 13: 55–72.
6.
SungJHKamCShulerML. A microfluidic device for a pharmacokinetic-pharmacodynamic (PK-PD) model on a chip. Lab Chip2010; 10: 446–55.
7.
OleagaCBernabiniCSmithASSrinivasanBJacksonMMcLambWPlattVBridgesRCaiYSanthanamNBerryBNajjarSAkandaNGuoXMartinCEkmanGEschMBLangerJOuedraogoGCotovioJBretonLShulerMLHickmanJJ. Multi-Organ toxicity demonstration in a functional human in vitro system composed of four organs. Sci Rep2016; 6: 20030–20030.
8.
MillerPGShulerML. Design and demonstration of a pumpless 14 compartment microphysiological system. Biotechnol Bioeng2016; 113: 2213–27.
9.
EschMBUenoHApplegateDRShulerML. Modular, pumpless body-on-a-chip platform for the co-culture of GI tract epithelium and 3D primary liver tissue. Lab Chip2016; 16: 2719–29.
10.
Lee H, Kim DS, Ha SG, Choi I, Lee JM, Sung JH. A pumpless multi-organ-on-a-chip (MOC) combined with a pharmacokinetic-pharmacodynamic (PK-PD) model. Biotechnol Bioeng 2017;114:432–43.
11.
NatarajanAMolnarPSieverdesKJamshidiAHickmanJJ. Microelectrode array recordings of cardiac action potentials as a high throughput method to evaluate pesticide toxicity. Toxicol In Vitro2006; 20: 375–81.
12.
Wang YI, Abaci HE, Shuler ML. Microfluidic blood–brain barrier model provides in vivo-like barrier properties for drug permeability screening. Biotechnol Bioeng 2017;114:184–94.
13.
AbaciHEGledhillKGuoZChristianoAMShulerML. Pumpless microfluidic platform for drug testing on human skin equivalents. Lab Chip2015; 15: 882–8.
14.
ChenHJSunJHuangZHouHArcillaMRakhilinNJoeDJChoiJGadamsettyPMilsomJNandakumarGLongmanRZhouXKEdwardsRChenJChenKYBuPWangLXuYMunroeRAbratteCMillerADGümüşZHShulerMNishimuraNEdelmannWShenXLipkinSM. Comprehensive models of human primary and metastatic colorectal tumors in immunodeficient and immunocompetent mice by chemokine targeting. Nat Biotechnol2015; 33: 656–60.
15.
EschMBProtJ-MWangYIMillerPLlamas-VidalesJRNaughtonBAApplegateDRShulerML. Multi-cellular 3D human primary liver cell culture elevates metabolic activity under fluidic flow. Lab Chip2015; 15: 2269–77.
16.
GuoXAyalaJEGonzalezMStancescuMLambertSHickmanJJ. Tissue engineering the monosynaptic circuit of the stretch reflex arc with co-culture of embryonic motoneurons and proprioceptive sensory neurons. Biomaterials2012; 33: 5723–31.
17.
GuoXGonzalezMStancescuMVandenburghHHHickmanJJ. Neuromuscular junction formation between human stem cell-derived motoneurons and human skeletal muscle in a defined system. Biomaterials2011; 32: 9602–11.
18.
SungJHSrinivasanBEschMBMcLambWTBernabiniCShulerMLHickmanJJ. Using physiologically-based pharmacokinetic-guided “body-on-a-chip” systems to predict mammalian response to drug and chemical exposure. Exp Biol Med2014; 239: 1225–39.
19.
HynesRO. The extracellular matrix: not just pretty fibrils. Science2009; 326: 1216–9.
20.
BerryBJAkandaNSmithASLongCJSchnepperMTGuoXHickmanJJ. Morphological and functional characterization of human induced pluripotent stem cell-derived neurons (iCell Neurons) in defined culture systems. Biotechnol Prog2015; 31: 1613–22.
21.
SchaffnerAEBarkerJLStengerDAHickmanJJ. Investigation of the factors necessary for growth of hippocampal neurons in a defined system. J Neurosci Methods1995; 62: 111–9.
22.
SmithASLongCJPirozziKNajjarSMcAleerCVandenburghHHHickmanJJ. A multiplexed chip-based assay system for investigating the functional development of human skeletal myotubes in vitro. J Biotechnol2014; 185: 15–8.
23.
StancescuMMolnarPMcAleerCWMcLambWLongCJOleagaCProtJMHickmanJJ. A phenotypic in vitro model for the main determinants of human whole heart function. Biomaterials2015; 60: 20–30.
24.
GuoXJoheKMolnarPDavisHHickmanJ. Characterization of a human fetal spinal cord stem cell line, NSI-566RSC, and its induction to functional motoneurons. J Tissue Eng Regen Med2010; 4: 181–93.
25.
RumseyJWDasMBhalkikarAStancescuMHickmanJJ. Tissue engineering the mechanosensory circuit of the stretch reflex arc: sensory neuron innervation of intrafusal muscle fibers. Biomaterials2010; 31: 8218–27.
26.
DavisHGonzalezMStancescuMLoveRHickmanJJLambertS. A phenotypic culture system for the molecular analysis of CNS myelination in the spinal cord. Biomaterials2014; 35: 8840–5.
27.
WilsonKAFinchCAAndersonPVollmerFHickmanJJ. Whispering gallery mode biosensor quantification of fibronectin adsorption kinetics onto alkylsilane monolayers and interpretation of resultant cellular response. Biomaterials2012; 33: 225–36.
28.
SpargoBJTestoffMANielsenTBStengerDAHickmanJJRudolphAS. Spatially controlled adhesion, spreading, and differentiation of endothelial cells on self-assembled molecular monolayers. Proc Nat Acad Sci1994; 91: 11070–4.
29.
StengerDAPikeCJHickmanJJCotmanCW. Surface determinants of neuronal survival and growth on self-assembled monolayers in culture. Brain Res1993; 630: 136–47.
30.
HickmanJJBhatiaSKQuongJNShoenPStengerDAPikeCJCotmanCW. Rational pattern design for in vitro cellular networks using surface photochemistry. J Vacuum Sci Technol A1994; 12: 607–607.
31.
WilsonKDasMWahlKJColtonRJHickmanJ. Measurement of contractile stress generated by cultured rat muscle on silicon cantilevers for toxin detection and muscle performance enhancement. PLoS One2010; 5: e11042–e11042.
32.
WilsonKMolnarPHickmanJ. Integration of functional myotubes with a Bio-MEMS device for non-invasive interrogation. Lab Chip2007; 7: 920–2.
33.
MolnarPWangWSNatarajanARumseyJWHickmanJJ. Photolithographic patterning of C2C12 myotubes using vitronectin as growth substrate in serum-free medium. Biotechnol Prog2007; 23: 265–8.
34.
DasMGregoryCAMolnarPRiedelLMWilsonKHickmanJJ. A defined system to allow skeletal muscle differentiation and subsequent integration with silicon microstructures. Biomaterials2006; 27: 4374–80.
35.
DasMRumseyJWBhargavaNGregoryCRiedelLKangJFHickmanJJ. Developing a novel serum-free cell culture model of skeletal muscle differentiation by systematically studying the role of different growth factors in myotube formation. In Vitro Cell Dev Biol Anim2009; 45: 378–87.
36.
DasMRumseyJWBhargavaNStancescuMHickmanJJ. Skeletal muscle tissue engineering: a maturation model promoting long-term survival of myotubes, structural development of the excitation-contraction coupling apparatus and neonatal myosin heavy chain expression. Biomaterials2009; 30: 5392–402.
37.
DasMRumseyJWBhargavaNStancescuMHickmanJJ. A defined long-term in vitro tissue engineered model of neuromuscular junctions. Biomaterials2010; 31: 4880–8.
38.
DasMWilsonKMolnarPHickmanJJ. Differentiation of skeletal muscle and integration of myotubes with silicon microstructures using serum-free medium and a synthetic silane substrate. Nat Protoc2007; 2: 1795–801.
39.
PirozziKLLongCJMcAleerCWSmithASHickmanJJ. Correlation of embryonic skeletal muscle myotube physical characteristics with contractile force generation on an atomic force microscope-based bio-microelectromechanical systems device. Appl Phys Lett2013; 103: 83108–83108.
40.
McAleerCWSmithASNajjarSPirozziKLongCJHickmanJJ. Mechanistic investigation of adult myotube response to exercise and drug treatment in vitro using a multiplexed functional assay system. J Appl Physiol (1985)2014; 117: 1398–405.
41.
NatarajanAChunCHickmanJJMolnarP. Growth and electrophysiological properties of rat embryonic cardiomyocytes on hydroxyl- and carboxyl-modified surfaces. J Biomater Sci Polym Ed2008; 19: 1319–31.
42.
SmithASLongCJPirozziKHickmanJJ. A functional system for high-content screening of neuromuscular junctions. Technology2013; 1: 37–48.
43.
NatarajanAStancescuMDhirVArmstrongCSommerhageFHickmanJJMolnarP. Patterned cardiomyocytes on microelectrode arrays as a functional, high information content drug screening platform. Biomaterials2011; 32: 4267–74.
44.
WilsonKStancescuMDasMRumseyJHickmanJ. Direct patterning of coplanar polyethylene glycol alkylsilane monolayers by deep-ultraviolet photolithography as a general method for high fidelity, long-term cell patterning and culture. J Vacuum Sci Technol B2011; 29: 0210201–10–0210201–10.
45.
EdwardsDStancescuMMolnarPHickmanJJ. Two cell circuits of oriented adult hippocampal neurons on self-assembled monolayers for use in the study of neuronal communication in a defined system. ACS Chem Neurosci2013; 4: 1174–82.
SinAChinKCJamilMFKostovYRaoGShulerML. The design and fabrication of three-chamber microscale cell culture analog devices with integrated dissolved oxygen sensors. Biotechnol Prog2004; 20: 338–45.
48.
MahlerGJEschMBGlahnRPShulerML. Characterization of a gastrointestinal tract microscale cell culture analog used to predict drug toxicity. Biotechnol Bioeng2009; 104: 193–205.
49.
WagnerIMaterneEMBrinckerSSussbierUFradrichCBusekMSonntagFSakharovDATrushkinEVTonevitskyAGLausterRMarxU. A dynamic multi-organ-chip for long-term cultivation and substance testing proven by 3D human liver and skin tissue co-culture. Lab Chip2013; 13: 3538–47.
50.
MaschmeyerILorenzAKSchimekKHasenbergTRammeAPHubnerJLindnerMDrewellCBauerSThomasASamboNSSonntagFLausterRMarxU. A four-organ-chip for interconnected long-term co-culture of human intestine, liver, skin and kidney equivalents. Lab Chip2015; 15: 2688–99.
51.
AnFQuYLuoYFangNLiuYGaoZZhaoWLinB. A laminated microfluidic device for comprehensive preclinical testing in the drug ADME process. Sci Rep2016; 6: 25022–25022.
52.
SungJHEschMBProtJMLongCJSmithAHickmanJJShulerML. Microfabricated mammalian organ systems and their integration into models of whole animals and humans. Lab Chip2013; 13: 1201–12.
53.
SakolishCMEschMBHickmanJJShulerMLMahlerGJ. Modeling barrier tissues in vitro: methods, achievements, and challenges. EBioMedicine2016; 5: 30–9.
54.
SrinivasanBKolliAREschMBAbaciHEShulerMLHickmanJJ. TEER measurement techniques for in vitro barrier model systems. J Lab Autom2015; 20: 107–26.
55.
CaliCell instruction manual: World Precision Instruments, 2014.
56.
BoothRKimH. Characterization of a microfluidic in vitro model of the blood–brain barrier (muBBB). Lab Chip2012; 12: 1784–92.
57.
BergerDRWareBRDavidsonMDAllsupSRKhetaniSR. Enhancing the functional maturity of induced pluripotent stem cell-derived human hepatocytes by controlled presentation of cell–cell interactions in vitro. Hepatology2015; 61: 1370–81.
58.
CameronKTanRSchmidt-HeckWCamposGLyallMJWangYLucendo-VillarinBSzkolnickaDBatesNKimberSJHengstlerJGGodoyPForbesSJHayDC. Recombinant laminins drive the differentiation and self-organization of hESC-derived hepatocytes. Stem Cell Rep2015; 5: 1250–62.
59.
ChoiKPfundWPAndersenMEThomasRSClewellHJLeCluyseEL. Development of 3D dynamic flow model of human liver and its application to prediction of metabolic clearance of 7-ethoxycoumarin. Tissue Eng Part C Methods2014; 20: 641–51.
60.
FerriniJBPichardLDomergueJMaurelP. Long-term primary cultures of adult human hepatocytes. Chem Biol Interact1997; 107: 31–45.
61.
WareBRBergerDRKhetaniSR. Prediction of drug-induced liver injury in micropatterned co-cultures containing iPSC-derived human hepatocytes. Toxicol Sci2015; 145: 252–62.
62.
SungJHChoiJRKimDShulerML. Fluorescence optical detection in situ for real-time monitoring of cytochrome P450 enzymatic activity of liver cells in multiple microfluidic devices. Biotechnol Bioeng2009; 104: 516–25.
63.
AbaciHShulerM. Human-on-a-chip design strategies and principles for physiologically based pharmocokinetics/pharmacodynamics modeling. Integr Biol2015; 7: 383–91.
64.
ToepkeMWBeebeDJ. PDMS absorption of small molecules and consequences in microfluidic applications. Lab Chip2006; 6: 1484–6.
65.
WangJDDouvilleNJTakayamaSElSayedM. Quantitative analysis of molecular absorption into PDMS microfluidic channels. Ann Biomed Eng2012; 40: 1862–73.
66.
Gomez-SjobergRLeyratAAHousemanBTShokatKQuakeSR. Biocompatibility and reduced drug absorption of sol-gel-treated poly(dimethyl siloxane) for microfluidic cell culture applications. Anal Chem2010; 82: 8954–60.
67.
ZhengFFuFChengYWangCZhaoYGuZ. Organ-on-a-chip systems: microengineering to biomimic living systems. Small2016; 12: 2253–82.
68.
PardridgeWM. The blood–brain barrier: bottleneck in brain drug development. NeuroRx2005; 2: 3–14.
69.
WolffAAntfolkMBrodinBTenjeM. In vitro blood-brain barrier models – an overview of established models and new microfluidic approaches. J Pharm Sci2015; 104: 2727–46.
70.
BrownJAPensabeneVMarkovDAAllwardtVNeelyMDShiMBrittCMHoilettOSYangQBrewerBMSamsonPCMcCawleyLJMayJMWebbDJLiDBowmanABReisererRSWikswoJP. Recreating blood–brain barrier physiology and structure on chip: a novel neurovascular microfluidic bioreactor. Biomicrofluidics2015; 9: 054124–054124.
71.
MahlerGJShulerMLGlahnRP. Characterization of Caco-2 and HT29-MTX cocultures in an in vitro digestion/cell culture model used to predict iron bioavailability. J Nutr Biochem2009; 20: 494–502.
72.
MahlerGJEschMBTakoESouthardTLArcherSDGlahnRPShulerML. Oral exposure to polystyrene nanoparticles affects iron absorption. Nat Nanotechnol2012; 7: 264–71.
EschMBSungJHYangJYuCYuJMarchJCShulerML. On chip porous polymer membranes for integration of gastrointestinal tract epithelium with microfluidic ‘body-on-a-chip' devices. Biomed Microdevices2012; 14: 895–906.
75.
ChenHJWeiZSunJBhattacharyaASavageDJSerdaRMackeyevYCurleySABuPWangLChenSCohen-GouldLHuangEShenXLipkinSMCopelandNGJenkinsNAShulerML. A recellularized human colon model identifies cancer driver genes. Nat Biotechnol2016; 34: 845–51.
76.
SiramshettyVBNickelJOmieczynskiCGohlkeBODrwalMNPreissnerR. WITHDRAWN – a resource for withdrawn and discontinued drugs. Nucleic Acids Res2016; 44: D1080–6.
77.
GuoXGreeneKAkandaNSmithAStancescuMLambertSVandenburghHHickmanJ. In vitro differentiation of functional human skeletal myotubes in a defined system. Biomater Sci2014; 2: 131–8.
78.
HuangZJLuoL. NEUROSCIENCE. It takes the world to understand the brain. Science2015; 350: 42–4.
79.
EdwardsDDasMMolnarPHickmanJJ. Addition of glutamate to serum-free culture promotes recovery of electrical activity in adult hippocampal neurons in vitro. J Neurosci Methods2010; 190: 155–63.
80.
GuoXSpradlingSStancescuMLambertSHickmanJJ. Derivation of sensory neurons and neural crest stem cells from human neural progenitor hNP1. Biomaterials2013; 34: 4418–27.
81.
DavisHGonzalezMBhargavaNStancescuMHickmanJJLambertS. Rat cortical oligodendrocyte-embryonic motoneuron co-culture: an in vitro axon-oligodendrocyte interaction model. J Biomater Tissue Eng2012; 2: 206–14.
82.
RumseyJWMcAleerCDasMBhalkikarAWilsonKStancescuMLambertSHickmanJJ. Myelination and node of Ranvier formation on sensory neurons in a defined in vitro system. In Vitro Cell Dev Biol Anim2013; 49: 608–18.
83.
GuoXDasMRumseyJGonzalezMStancescuMHickmanJ. Neuromuscular junction formation between human stem-cell-derived motoneurons and rat skeletal muscle in a defined system. Tissue Eng Part C Methods2010; 16: 1347–55.
84.
SmithASLongCJPirozziKHickmanJJ. A functional system for high-content screening of neuromuscular junctions in vitro. Technology2013; 1: 37–48.
85.
Guo X, Sommerhage F, McAleer C, Martin C, Long C, Wang Y, Santhanam N, Colon A, Sancho CO, Hickman J. In vitro modeling of nervous system: engineering of the reflex arc. In: Zhang LG, Kaplan DL (eds) Neural engineering: from advanced biomaterials to 3D fabrication techniques. Cham: Springer International Publishing, 2016, pp.261–98.
86.
WilkeRALinDWRodenDMWatkinsPBFlockhartDZinehIGiacominiKMKraussRM. Identifying genetic risk factors for serious adverse drug reactions: current progress and challenges. Nat Rev Drug Discov2007; 6: 904–16.
87.
GodoyPHewittNJAlbrechtUAndersenMEAnsariNBhattacharyaSBodeJGBolleynJBornerCBottgerJBraeuningABudinskyRABurkhardtBCameronNRCamussiGChoCSChoiYJCraig RowlandsJDahmenUDammGDirschODonatoMTDongJDooleySDrasdoDEakinsRFerreiraKSFonsatoVFraczekJGebhardtRGibsonAGlanemannMGoldringCEGomez-LechonMJGroothuisGMGustavssonLGuyotCHallifaxDHammadSHaywardAHaussingerDHellerbrandCHewittPHoehmeSHolzhutterHGHoustonJBHrachJItoKJaeschkeHKeitelVKelmJMKevin ParkBKordesCKullak-UblickGALeCluyseELLuPLuebke-WheelerJLutzAMaltmanDJMatz-SojaMMcMullenPMerfortIMessnerSMeyerCMwinyiJNaisbittDJNusslerAKOlingaPPampaloniFPiJPlutaLPrzyborskiSARamachandranARogiersVRoweCSchelcherCSchmichKSchwarzMSinghBStelzerEHStiegerBStoberRSugiyamaYTettaCThaslerWEVanhaeckeTVinkenMWeissTSWideraAWoodsCGXuJJYarboroughKMHengstlerJG. Recent advances in 2D and 3D in vitro systems using primary hepatocytes, alternative hepatocyte sources and non-parenchymal liver cells and their use in investigating mechanisms of hepatotoxicity, cell signaling and ADME. Arch Toxicol2013; 87: 1315–530.
88.
EbrahimkhaniMRNeimanJARaredonMSHughesDJGriffithLG. Bioreactor technologies to support liver function in vitro. Adv Drug Deliv Rev2014; 69–70: 132–57.
89.
LiAPBodeCSakaiY. 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. Chem Biol Interact2004; 150: 129–36.
90.
WikswoJPCurtisELEagletonZEEvansBCKoleAHofmeisterLHMatloffWJ. Scaling and systems biology for integrating multiple organs-on-a-chip. Lab Chip2013; 13: 3496–511.
91.
MoraesCLabuzJMLeungBMInoueMChunTHTakayamaS. On being the right size: scaling effects in designing a human-on-a-chip. Integr Biol2013; 5: 1149–61.
92.
VozziFMazzeiDVinciBVozziGSbranaTRicottiLForgioneNAhluwaliaA. A flexible bioreactor system for constructing in vitro tissue and organ models. Biotechnol Bioeng2011; 108: 2129–40.
93.
EschMBMahlerGJStokolTShulerML. Body-on-a-chip simulation with gastrointestinal tract and liver tissues suggests that ingested nanoparticles have the potential to cause liver injury. Lab Chip2014; 14: 3081–92.
