Table 1.

Summary of selected microfluidic systems used in breast cancer research.

Reference	Cell Used	Culture Type	Field of Investigation	Device Properties	Findings
Cheng [19]	Acellular	2D	Cancer formation	Extract cell-free DNA from plasma to detect BRCA1 and BRCA2 mutations.	Successful detection of BRCA1/2 mutations with a minimum detectable number of copies of 20,000.
				Uses four distinct primers in parallel to provide point of care risk assessment.
				Dimensions not described, no ECM used, utilizes pressure differential pumping.
Pradhan [20]	MCF7 and MDA-MB-231 cancer, hBTEC endothelial, BJ5ta fibroblast	3D	Cancer formation	Two distinct chips with normo- and pathophysiologic vascular layout, respectively, used to assess anti-cancer drug delivery and tumor reaction to treatment.	Cells found to elongate and align along flow, dependent on the cell line.
				Model of cancer–stromal–endothelial interactions within a pillar-filled tumor region adjacent to vessels.	MCF7 found to have significant resistance to anti-cancer drug paclitaxel in low-perfusion chip design.
				100 μm channel width, PEG–fibrinogen hydrogel matrix, high perfusion layout experiences 40–50 s−1 shear rate, low perfusion chip experiences 10–20 s−1 shear rate.	Both MCF7 and MDA-MB-231 found to have significant resistance to anti-cancer drug doxorubicin in low-perfusion chip design.
Ayuso [21]	MCF10A cancer, HMF fibroblasts	3D	Cancer formation	Ductal carcinoma in situ in a central vessel surrounded by stroma-filled matrix between two empty channels for perfusion, metabolism, mobility, and gene expression investigation.	Hypoxia-activated Tirapazamine selectively destroys DCIS cells.
				Breast cancer cells, fibroblasts, and hydrogel model of luminal mammary duct.	Hypoxia-activated glycolysis transcriptome upregulation.
				Dimensions not described, collagen hydrogel, static conditions with daily media change.
Funamoto [22]	MDA-MB-231 cancer	3D	Cancer formation	Five-channel chip with central tumor model, surrounded by two media perfusion channels, which are, in turn, surrounded by two gas perfusion channels.	Hypoxia found to significantly increase cancer mobility.
				Breast cancer cells in 3D culture with controlled oxygen perfusion to investigate hypoxia effects.
				1.3 mm tumor channel width, 0.5 mm media and gas channel widths, collagen I hydrogel, 30 μL/h flow.
Tang [23]	MDA-MB-231 and MCF-7 cancer, primary human-breast-tumor-associated endothelial cells	3D	Cancer formation	Microfluidic chip modeled on 2D projection of tumor vasculature to investigate enhanced permeability and retention (EPR) found in tumors.	TNF-α found to significantly increase the permeability of endothelial cells.
				Breast cancer and endothelial cell biomimetic tumor microenvironment.	Tumor cell co-culture significantly increases the permeability of endothelial cells.
				100 μm channel width, fibronectin ECM, 60 uL/h or 0–90 s−1 shear rate.	Liposome extravasation through endothelial cells found to significantly increase during tumor co-culture.
Nashimoto [24]	MCF-7 and GFP MDA-MB-231 cancer, RFP HUVECs, normal human lung fibroblasts, SW620 with luciferase, Hepg2	Spheroids	Cancer formation	2 or 3 cell type co-culture spheroids of various cancers in 96-well plate transferred into microfluidic chip for angiogenesis.	Fibroblast co-culture induced angiogenic sprouts.
				1 mm width culture channel, fibrin-collagen matrix, 30 μL/h.	Flow reduces anti-cancer drug paclitaxel efficacy and leads to less necrotic tumors.
Uliana [32]	Acellular	2D	Cancer formation	Disposable microfluidic electrochemical array device to detect estrogen receptor alpha (ERα).	USD 0.20 cost of manufacture.
				Highly decorated magnetic particles and protein–DNA interaction detected using electrodes to quantify cancer signals.	Ultralow detection limit of 10.0 fg mL−1 for the determination of ERα in calf serum.
				3 mm wide and 200 mm long, no ECM, 6000 μL/h.	Good recoveries for detection of the biomarker in MCF-7 cell lysate.
Moon [44]	MCF-7, MDA-MB-231, and SUM-159PT cancer	3D	Cancer formation	Hydrogel tube seeded with breast cancers to quantify breast cancer subtype motility.	SUM-159PT found to be most invasive cell line.
				500 μm diameter, collagen I hydrogel, static conditions.	CD24 expression was elevated in 3D compared with 2D cultures.
Yong [45]	SUM149, HCC1937, MDA-MB-231, and BT549 cancer	3D	Cancer formation	A hydrogel channel with two perfused vessels, one seeded with breast cancer cells to quantify the directional migration of cancer cell lines.	MDA-MB-231 found to be most invasive.
				200 μm diameter, collagen hydrogel, static conditions.	305 genes identified as altered during invasion.
Wang [251]	SK-BR-3 cancer, HEK293FT cells, human T cells, Jurkat cells	Suspended cells	Cancer formation	A pair of chips, one that generates droplets containing cells and antibodies or lentivirus, another that electrically sorts droplets based on fluorescence.	CD40 antibody developed.
				Continuous flow droplet-based lentivirus transduction and antibody screening.	Active anti-Her2 × anti-CD3 BiTE antibodies developed using antibody library.
				40 μm deep channels, no ECM used, 600–1800 μL/h.
Sung [46]	MCF-DCIS.com cancer cells, GFP Human mammary fibroblasts	3D	Invasion	Mammary endothelial and fibroblast cells co-cultured in Y-shaped compartmentalized model.	Co-culture promotes invasion, diminishing as distance between cell types increases
				Second harmonic collagen imaging provides an index of transition to invasion.	Soluble factors shown to begin migration, while cell–cell contact between cancer and stromal cells completed the transition to invasion.
				1.5 mm width, Matrigel and collagen I matrix, static culture.
Lam [47]	RFP MDA-MB-231, skin fibroblasts	3D	Invasion	Two parallel 3D models with breast cancer and fibroblast co-cultures to study the role of acidity in the transition to invasion.	Calcium bicarbonate buffer nanoparticles inhibit acid-induced invasion.
				Channels between 200–1000 μm wide, fibrin hydrogel, 36 μL/h	Buffering selectively inhibited the growth of the MDA-MB-231.
Toh [48]	MX-1, MCF7 cancer	3D	Invasion	Tumor cells encased in collagen exposed to flowing chemoattractant signals on either side.	Cells found to exhibit collective, amoeboid, and mesenchymal-like motility.
				Real-time monitoring of cell extravasation	Distinct populations of collagen-penetrating invasive cancer cells and collagen-avoidant migratory cancer cells identified.
				1 cm long, 600 μm wide, and 100 μm tall, positively charged collagen and a negatively charged HEMA-MMA-MAA terpolymer matrix, 30 μL/h.
Yankaskas [49]	ZR75-1, MDA-MB-468, MDA-MB-436, Hs578t, BT-549, and MDA-MB-231 cancer cells, MCF-10A, T47, and human mammary epithelial cells, and HCC1428	3D	Invasion	Continuous flow of breast cancer cells near parallel apertures through which chemoattractants diffuse to induce migration.	Identified motility- and survival-related genes.
				Quantifies extravasation potential, abundance, and proliferative index of breast cancer cell subtypes.	MDA-MB-231 found to be most migratory.
				Width of 20 µm and a height of 10 µm, collagen matrix, flow not specified.	Self-sorted migratory cells found to be significantly more proliferative and preferentially localize to bone in comparison with non-sorted cells.
Gioiella [50]	MCF7 cancer cells, normal and cancer activated fibroblasts	3D	Invasion	Breast cancer tumor model using epithelial and stromal cell co-culture.	Hyaluronic acid and fibronectin overexpression are associated with invasion
				Macromolecule and collagen production quantified.	Fibroblasts significantly lose diffusivity when activated by nearby tumor cells.
				370 μm long, 780 μm wide, 300 μm deep, other channel is 1200 μm long, 1370 μm wide, 300 μm deep, crosslinked type A porcine-derived gelatin matrix, 180 µL/h.
Mosadegh [51]	A549	3D	Invasion	Stacked paper seeded with hydrogel and cancer cells, used to examine hypoxia-induced migration.	Oxygen acts as a chemoattractant for cancer cells.
				No channels, Matrigel matrix, no perfusion.	Distinct rates of oxygen chemotaxis between cell lines.
Lugo-Cintron [53]	MDA-MB-231 cancer cells, normal and cancer activated fibroblasts	3D	Invasion	Large channel containing fibroblasts in microenvironment with a central channel seeded with breast cancer cells.	Fibronectin-rich matrix associated with increased migration.
				3D breast cancer and fibroblast tumor model to visualize invasion.	MMP secretion associated with increased migration.
				400 μm wide central channel, rat-tail collagen type I, fibronectin, and fibrin matrix, static culture.
Buchanan [72]	MDA-MB-231 cancer cells, telomerase immortalized microvascular endothelial (TIME) cells	3D	Intravasation	Microchannel embedded within a collagen hydrogel as a vessel model with shear quantified using microparticle image velocimetry.	Tumor cells significantly increase the expression of proangiogenic genes in response to co-culture with endothelial cells under low flow conditions.
				Microfluidic tumor vascular model for co-culture of tumor and endothelial cells under varying flow shear stress conditions	Endothelial cells develop a confluent endothelium on the microchannel lumen that maintains integrity under physiological flow shear stresses.
				850 μm diameter, Collagen 1 matrix, 180 μL/h.
Choi [73]	Human primary mammary fibroblasts, HMT-3522 cells	Spheroids, 3D	Intravasation	Intravasation model using a two-channel stroma and endothelial co-culture device later seeded with human mammary ductal epithelial cells and mammary fibroblasts co-culture breast tumor spheroids.	Paclitaxel anti-cancer drug shown to inhibit progression through cytotoxicity.
				Visualization of attachment and intravasation.
				1 mm wide, 3 mm long, and 200 μm tall, collagen hydrogel, 60 μL/h.
Nagaraju [76]	MDA-MB-231 and MCF7 cancer cells, HUVECs	3D	Intravasation	Chip with distinct tumor, stroma, and vascular channels to model invasion and intravasation into media-filled vascular channel.	Endothelial cells significantly increase the migration of tumor cells.
				200 μm wide channel, collagen matrix, flow rate not specified.	Tumor signaling significantly reduces vessel diameter and increases endothelial permeability.
					Absence of endothelial cells significantly alters the secretion of ANG-2 and angiogenin.
Zervantonakis [71]	MDA231 cancer cells, HT1080 fibroblasts	3D	Intravasation	Three-channel tumor model with an endothelial channel and a tumor channel separated by a cell-free 3D ECM channel to model intravasation towards endothelial cells.	Macrophage-secreted tumor necrosis factor alpha significantly increases intravasation rate.
				500 μm wide, 20 mm in length, and 120 μm in height, collagen matrix, flow not specified.	Endothelium provides a barrier to intravasation, regulated by tumor microenvironment factors.
Cui [75]	MDA-MB-231 cancer cells, primary human vascular endothelial cells	3D	Intravasation	32 independent cell collection microchambers with endothelial layer for characterization of trans-endothelial migration.	Migratory cancer cells significantly alter Palladin expression, F-actin orientation, and cell aspect ratio.
				4 mm by 4 mm chip, poly-D-lysine and fibronectin matrix, 1200 µL/h.	Different cancer lines show significantly distinct sensitivity to shear stress impact on trans-endothelial migration
Shirure [77]	MDA-MB-231 and MCF-7 cancer cells, Endothelial colony forming cell-derived endothelial cells, normal human lung fibroblasts, colorectal cancer cell line Caco-2	3D and Spheroid	Intravasation	Two tumor model chambers separated by one fibroblast and endothelial cell vascular model chamber to model arterial capillary intravasation.	VEGF and TGFβ significantly elevated during intravasation and migration.
				355 μm wide, fibrin matrix, 10 mm H2O pressure head.	Tumor cells expressing mesenchymal-like transcriptome invade into vascular chamber significantly more efficiently.
Jiang [146]	Lung, breast, and melanoma cancer blood samples	Suspended cells	Circulating signals and cells	Chip using deterministic lateral displacement to enrich platelet-CTC aggregates in conjugation with platelet antibodies for isolation.	60% reliable isolation of breast cancer CTC clusters.
				24 parallel channels that are 150 μm in depth, no ECM, 1000 μL/h.
Au [147]	Primary breast cancer CTCs and CTC clusters	Suspended cells	Circulating signals and cells	Two-stage continuous isolation of CTC clusters using asymmetry induced rotation.	2–100+ cells recovered from whole blood.
				99% recovery of large clusters, cell viability over 87%.
Deliorman [149]	PC3 human prostate cancer cell line	Suspended cells	Circulating signals and cells	Surface-bound antibodies for EpCAM, PSA, and PSMA to isolate CTCs for AFM measurements.	CTCs from metastatic cancer have decreased elasticity and increased deformability compared with tumor cells.
				900 μm wide, 85 μm deep, and 48 mm long, no ECM, 1200 μL/h.	Fewer multiple adhesion events in CTCs compared with tumor cells.
Sarioglu [158]	MDA-MB-231	Suspended cells	Circulating signals and cells	Detection of CTCs independent of tumor-specific markers using bifurcating traps under low-shear conditions.	30–40% successful detection of CTC clusters.
				4096 parallel 60 μm wide traps, no ECM, 2.5 mL/h.	RNA sequencing identifies macrophages within CTC clusters are tissue-derived from the primary site.
de Oliveira [163]	Acellular	Suspended cells	Circulating signals and cells	CTC detection using double-layer capillary capacitors to quantify CA 15-3.	5µL sample volume and 92.0 μU mL−1 detection limit.
				Antibody-anchored magnetic bead capture of CTCs for analysis.	USD 0.97 cost of manufacture.
				800 μm outer diameter, 545 μm inner diameter, 200 μm electrode gap, no ECM, no flow.
Marrella [159]	MDA-MB-231	Suspended cells	Circulating signals and cells	Multi-channel device simultaneously analyze correlation between shear stress and CTC cluster behavior.	Higher values of wall shear stress significantly correlated with decreased CTC viability to metastasize.
				1 mm wide and 120 mm long, no ECM, 0–20 dyne/cm2.	High shear stress significantly disaggregates CTC clusters within 6 h.
Regmi [160]	MDA-MB-231 and UACC-893 cancer cells, lung cancer A549, ovarian cancer 2008, leukemia K562 cells	Suspended cells	Circulating signals and cells	Microfluidic circulatory system to produce relevant shear stress levels on CTCs and CTC clusters to investigate disaggregation under shear.	60 dynes/cm2 during exercise leads to necrosis or apoptosis in 90% of CTCs after 4 h of circulation, significantly more than 15 dynes/cm2.
				1mm diameter tube, no ECM, 15–60 dyne/cm2.	High shear significantly reduces metastatic potential and drug resistance of breast cancer cells.
Uliana [32]	MCF-7 cells, a human breast cancer cell line	Suspended cells	Circulating signals and cells	Disposable microfluidic electrochemical arrays using electrode-bound DNA and antibody-conjugated magnetic beads to detect estrogen receptor alpha in plasma.	10.0 fg mL−1 detection limit.
				1.5 mm diameter electrode, no ECM, 6000 μL/h.	94.7–108% recovery of estrogen receptor alpha.
					USD 0.20 cost of manufacture.
Vaidyanathan [187]	BT-474 and MDA-MB-231 cancer cells, PC3 prostate cancer cells	Suspended cells	Circulating signals and cells	Multiplexed electrohydrodynamic detection of exosome targets in a chip.	Isolation of exosomal samples from HER2 and prostate-specific antigen.
				400 μm wide, 300 μm tall, 25 mm length, no ECM, 420 μL/h.	8300 exosomes/µL detection sensitivity.
Gao [191]	Acellular	Suspended cells	Circulating signals and cells	Multiple detections of miRNAs from multiple samples using three-segment hybridization detection in a microfluidic chip.	Successful detection of four breast cancer biomarker miRNAs.
				50 independent channels on a 25 by 75 mm2 chip, no ECM, flow not described.	Detection limit of 1 pM in 30 min.
Armbrecht [110]	MCF-7, SK-BR-3, and BR16 cancer cells, LM2 variant cell line	Suspended cells	Organotropism and extravasation	Integrated capture, isolation, and membrane analysis of CTCs in a chip.	95% isolation efficiency, granulocyte growth-stimulating factor detection efficiency at 1.5 ng/mL−1.
				30 independent 75 μm width chambers, no ECM, 12 μL/h.
Park [111]	MCF-7 and MDA-MB-231 cancer cells, HL-60 promyelocytic leukemia cell line	Suspended cells	Organotropism and extravasation	Inertial-force-assisted droplet generation using spirals to generate CTC clusters with known cell type ratios.	E-cadherin, VCAM-1, and mRNA expression characterized between cluster compositions.
				140 μm width and five-loop structure, no ECM, 1200 μL/h.
Riahi [113]	MDA-MB-231 cancer cells, HUVECs, human mammary epithelial cells, MCF7	Suspended cells, 3D	Organotropism and extravasation	Organ-specific extravasation of CTCs through an endothelial layer using localized chemokine gradients in a chip.	CXCL12 found to significantly increase extravasation.
				150 μm by 500 μm by 3 cm, Matrigel matrix, 50 μL/h.
Aleman Sarkal [114]	HUVECs, HCT-116 colorectal cancer, HEPG2, A549	Suspended cells, 3D	Organotropism and extravasation	Organ-specific extravasation of CTCs modeled using bioengineered 3D organoids of five different tissues.	HCT115 CRC cells preferentially extravasate into liver and lung cells, as seen in vivo.
				200 μm width, HA/gelatin matrix, 600 μL/h.
Song [115]	MDA-MB-231, MCF-10A, and MCF-7 cancer cells, HUVECs, normal human lung fibroblasts	Suspended cells, 3D	Organotropism and extravasation	Microvascular network model to quantify the extravasation of breast cell lines in distinct oxygen conditions.	HIF-1a confirmed through knockout and siRNA to significantly increase the transmigration capacity in breast cell lines and regulate apoptotic-related cellular processes.
				1 mm wide and 150 µm deep, fibrin matrix, static conditions.	In hypoxia, HIF-1a levels increased alongside changes in morphology and an increase in cancer viability and metastatic potential.
Marturano-Kruik [116]	GFP MDA-MB-231, Firefly MDA-MB-231, HUVECs, human bone marrow mesenchymal stem cells	3D	Organotropism and extravasation	Perfused bone perivascular niche on a chip to measure progression and drug resistance during metastasis.	Bone-marrow-derived mesenchymal stem cells shown to transition towards perivascular cell lineages and support capillary formation.
				Dimensions not shown, decellularized bone ECM matrix, 15 μL/h.	Interstitial flow within bone perivascular niche persists with low proliferation and high drug resistance.
Jeon [117]	MDA-MB-231 cancer, GFP HUVECs, human bone marrow mesenchymal stem cells	3D	Organotropism and extravasation.	Microfluidic bone, vascular, and myoblast model to analyze breast cancer organotropism.	Extravasation rate and permeability found to be significantly distinct between bone, myoblast, and unconditioned matrix models.
				1.3 mm wide and 200 μm deep, fibrin ECM, 120 μm/h.	A3 adenosine receptor disruption resulted in significantly higher extravasation rates to myoblast-containing matrix
Bersini [118]	GFP MDA-MB-231 cancer, RFP HUVECs, human bone marrow mesenchymal stem cells	3D	Organotropism and extravasation	Osteo-differentiated bone marrow-derived mesenchymal stem cells and endothelial cell bone microenvironment model of organotropism of breast cancer CTCs in a chip.	CTCs extravasated into the bone model 77.5% of the time at a distance of 50.8 µm, compared with 37.6% of the time at a distance of 31.8 µm into collagen control.
				Eight parallel 225 µm by 150 µm gel regions, collagen hydrogel ECM, static conditions.	Bone secreted signals CXCR2 and CXCL5 found to influence extravasation rate and travel distance.
Mei [119]	MDA-MB-231 cancer cells, HUVECs, MLO-Y4 osteocytes, RAW264.7 osteoclasts	3D	Organotropism and extravasation	Breast cancer, endothelial cell, and osteocyte-like cell model of bone extravasation using oscillatory shear in a chip.	3.71-fold increase in calcium response in 82.3% of osteocytes compared to continuous flow.
				1 mm by 200 μm static channel in addition to a 500 μm by 500 μm vascular model. Channel, collagen, and Matrigel matrix, 1.5 Pa wall shear stress.	Mechanical stimulation reduced extravasation distance 32.4% and frequency by 53.5%.
Xu [120]	MDA-MB-231 and M624 cells cancer, primary rat brain microvascular endothelial cells (BMECs), primary rat cerebral astrocytes	3D	Circulating signals and cells	Blood–brain barrier model in a chip.	Cancer cell and astrocyte interactions increase brain tumor migration between brain and vascular compartments.
				200 μm by 400 μm, collagen matrix, 0.1 dyne/cm2 and 60 μL/h.
Liu [270]	MCF-7 and MCF-7adm cancer	3D	Treatment	Five parallel gradient-generating networks on a chip using dam and weir structures for cell positioning and seeding to investigate anti-cancer drugs.	GSH levels of two breast cancer cell lines were reduced during arsenic trioxide treatment, resulting in increased chemotherapy sensitivity, and vice versa was found with N-acetyl cystine.
				2 mm by 1 mm by 30 μm, no ECM, 20 μL/h.
Parekh [275]	MDA-MB-231	Suspended particles	Treatment	Single breast cancer cell selection, drug loading, and fluorescence measurement on a chip.	Cyclosporine A found to greatly increase the cellular uptake of anti-cancer drugs by reducing drug efflux.
				30 mm by 30 mm chip, no ECM, no flow.
Sarkar [271]	MCF-7	Suspended particles	Treatment	Droplet docking microfluidic microarray to encapsulate single cells to investigate anti-cancer drug influx, efflux, and cytotoxicity.	Confirmation of previous findings and further classification of drug resistance between breast cancer cell lines.
				No dimensions given, no ECM, no flow.	Observed increased drug resistance during the homotypic fusion of cell-sensitive and resistant cell types within droplets.
Zhang [244]	2A4 hybridoma cells	Suspended particles	Treatment	Integrating chip for screening heavy and light chain combinations in antibodies using single-cell trapping, qPCR, and fluorescence to quantify specificity.	Anti-CD45 monoclonal antibodies identified using hybridoma cells.
				50 μm width, no ECM, 180 μL/h.
Cheng [233]	T47D and BT549 cancer, primary HUVECs	Spheroids	Treatment	Isolated tumor spheroids trapped next to endothelial cell vascular tissue model to analyze nanoparticle drug delivery systems.	The nanoparticle drug Ds-PEG-FA/DOX found to penetrate spheroids of BT549 cancer but not T47D.
				300 μm long, 200 μm wide, 100 μm deep, basement membrane extract matrix, flow rate not specified.
Qi [273]	MDA-MB-231 and MDA-MB-231/MDR	Subcellular and 2D	Treatment	Antibody-laden pillars for the specific capture of exosomes for isolation.	CD63-laden exosomes isolated at 70% efficiency.
				4 mm channel length, no ECM, 600 μL/h.	Drug content of isolated exosomes found to be significantly different between cell lines and between treatments.
Qiao [257]	A549 Human lung adeno-carcinoma, HEK293T human kidney cell line	Suspended particles	Treatment	Microfluidic encapsulation of oncolytic adenovirus and the BET bromodomain inhibitor JQ1 within PVA microgels for injection into tumors.	Found to extend persistence and accumulation of oncolytic adenovirus within tumors.
				Device dimensions not specified, no ECM, flow rate not specified.	PVA microgels inhibited PD-L1 expression to overcome immune suppression.
Ide [252]	BW5147 (H2k), JCRB9002, and BW5147	Suspended cells	Treatment	Lymph node and T cell–APC interaction model for cell collection and analysis.	Calcium ion flux fluorescent dye in T cells used as metric of activation during serial contact with APCs.
				20 μm well diameter, no ECM, 4200 μL/h.	T cell activation during contact with OVA 257–264 peptide presenting quantified APCs.
Lou [239]	Bone marrow cells	Suspended particles	Treatment	Tunable liposome formation in a microfluidic channel.	Cationic liposomes of 50–750 nm composed of combinations of tailored phospholipid ratios.
				Hydrodynamic focusing cartridges of 10 and 65 µm width, no ECM, 900 mL/h.	Macrophage liposome uptake is modulated by area and volume, and biodistribution in mice showed that <50 nm particles increase clearance rate.
Liu [240]	HeLa	3D	Treatment	Continuous flow co-precipitation of polymers to produce injectable “nanosystems”.	Nanosystem loaded with photosensitive zinc successfully localized to cancer cells to enable photodynamic therapy.
				200 μm wide and 45 μm deep, no ECM, 7200 μL/h.
Lee [259]	MRC5 fibroblasts, human lung cancer A549 cells	Spheroid	Treatment	Cancer, fibroblast, and endothelial spheroid model for oncolytic virus infection on a chip.	Cancer cells transfected substantially more than bystander cells, which is not observed in 2D cell culture
				500 μm spheroid chambers, collagen I matrix, 0.3 dyne/cm2.	HUVEC IFN-B secretion is delayed in 2D compared to the microfluidic model.
Terrell [248]	MDA-MB-231-HER2+ cancer, CTX-TNA2 rat astrocytes	3D	Treatment	Blood–brain barrier model in a chip to investigate monoclonal antibody localization.	Tailored antibody trastuzumab found to localize 3% to healthy brain and 5% to tumor model brain.
				200 μm width, fibronectin matrix, 60 μL/h.	Rate of uptake quantified as 2.7 × 103 to healthy brain and 1.28 × 104 to cancerous brain.
He [238]	Acellular	Suspended particles	Treatment	Hydrodynamic flow for self-assembly of amphiphilic nanoparticles capable of forming a “micelle”.	500 nm to 2 μm giant vesicles formed.
				Channel size not described, no ECM, 5400 μL/h.