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QUASI VIVO® GLOSSARY

Check out a compilation of information taken from publications and posters featuring Quasi Vivo®.

Mazzei et al (2010) - Designing a modular bioreactor with low shear stress and high flow rates for hepatocyte culture
Cell Types
Primary rat hepatocytes
Cell SourceAdult male rats (250-300g)
Flow Rate (µL/min)180; 60 -1000
Shear Pressure (Pa)4.34 x 10-^6; 6.5 x 10-^6 (at 300 µL/min)
Time in flow24 hours
Culture config/substrateCollagen sandwich
Seeding Density2x10^5 cells per cover slip
MediaFBS-free complete Williams' E
Media Volume10 mL
AssayViability, albumin ELISA
Vinci et al (A) (2011) - Liver detoxification gene expression and activity under flow
Cell Types
Primary human hepatocytes
Cell SourceIsolated from liver donors
Flow Rate (µL/min)250 - 500
Shear Pressure (Pa)5 x 10-^6 - 1 x 10-^5
Time in flow7 - 21 days
Culture config/substrateCollagen sandwich
Seeding Density~1.9x10^5 cells per cover slip
MediaLong term medium: Williams' E medium, 1.25 µM ethanolamine, 5 mg/L transferrin, 15.4 µM linoleic acid, 10 mg/L insulin, 0.1 µM selenium acid, 1 mg/mL glucagon, 0.1 µM dexamethasone, 5 µM cAMP, 10 IU/L prolactin, 2 g/L glucose, 50 µg/L epidermal growth factor, 20 µg/L liver growth factor, 0.25 mM sodium pyruvate, 2 mg/L ascorbic acid
Media Volume
AssayqRT-PCR, drug metabolism, albumin & urea secretion
Vozzi et al (2011) - In vitro tissue and organ modelling using connected bioreactors
Cell Types
HepG2 hepatocytes
Cell SourceHuman hepatoblastoma line
Flow Rate (µL/min)250
Shear Pressure (Pa)5 x 10-^6
Time in flow1 week; 48 hours
Culture config/substrateCollagen-coated PLGA scaffold, coated with alginate film
Seeding Density1x10^5 cells per scaffold
MediaConditioned endothelial cell growth medium containing 10% FBS
Media Volume15 mL
AssayGlucose, free fatty acid, glycerol, D-lactate, L-alanine and albumin concentration
Vinci et al (B) (2011) - Inter-organ crosstalk in a 3-way glucose and lipid metabolism model
Cell Types
HepG2 hepatocytes, primary human adipocytes (plus collagen matrix)
Cell SourceLaboratory of Molecular Hepatology, University of Padua; surgically isolated omental adipose tissue
Flow Rate (µL/min)250
Shear Pressure (Pa)1 x 10-^5
Time in flow15, 24 & 48 hours
Culture config/substrateCollagen-coated PLGA scaffold, coated with alginate film; nylon mesh
Seeding Density1x10^5 hepatocytes per scaffold
MediaConditioned endothelial cell growth medium containing 10% FBS
Media Volume15 mL
AssayGlucose, free fatty acid, glycerol, D-lactate, L-alanine and albumin concentration
Vozzi et al (2011) - Diclofenac dose-response in hepatocytes cultured in Quasi Vivo
Cell Types
Primary rat hepatocytes, gliosarcoma cells (connected culture)
Cell Source
Flow Rate (µL/min)100
Shear Pressure (Pa)2.9 x 10-^6
Time in flow24 hours
Culture config/substrateCoated in collagen gel
Seeding Density2x10^5 cells per well
MediaSerum-free Williams' medium
Media Volume
AssayAlmar blue vitality
Iori et al (2012) - Glycaemic challenge in a 3-way glucose and lipid metabolism model
Cell Types
HepG2 hepatocytes, primary human adipocytes (plus collagen matrix)
Cell SourceLaboratory of Molecular Hepatology, University of Padua; surgically isolated omental adipose tissue
Flow Rate (µL/min)250
Shear Pressure (Pa)1 x 10-^5
Time in flow15, 24 & 48 hours
Culture config/substrateCollagen-coated PLGA scaffold, coated with alginate film; nylon mesh
Seeding Density1x10^5 hepatocytes per scaffold
MediaEndothelial cell growth medium containing 10% FBS
Media VolumeEndothelial cell growth medium containing 10% FBS
AssayViability, morphology; glycerol, D-lactate, L-alanine (spectrophotometry); free fatty acids, glucose (enzyme assays); IL-6, albumin, E-selectin (ELISA)
InLiveTox consortium (2012) - Gut-liver-vascular in vitro model for assessing nanoparticle (NP) toxicity
Cell Types
Enterocytes (Caco-2), hepatocytes (C3A), endothelial cells (HUVECs)
Cell Source
Flow Rate (µL/min)
Shear Pressure (Pa)
Time in flow
Culture config/substrateUltrathin silicon nitride membrane with built-in TEER electrodes (for intestinal epithelial cell culture)
Seeding Density
MediaEMEM-GF (common medium for all 3 cell types)
Media Volume
AssayPhenotypic function endpoint assays: TEER (epithelial), vWF expression (endothelial), albumin expression (hepatocyte). Testing for aspects of NP toxicity: membrane damage (LDH, propidium iodide), cell death (alamarBlue), apoptosis (caspase activity, Fas-ligand), oxidative stress (GSH), inflammation (cytokine gene & protein expression by RT-PCR & Bioplex)
Giusti et al (2014) - Dual-flow intestinal barrier model
Cell Types
Caco-2/TC7 intestinal epithelial cells
Cell SourceSigma-Aldrich
Flow Rate (µL/min)Apical = 200, basal = 100
Shear Pressure (Pa)6 x 10-^4
Time in flow24 & 48 hours
Culture config/substrateMonolayer on polycarbonate membrane of QV600 insert
Seeding Density3x10^4 cells/cm2
MediaDulbecco’s modified medium (high glucose) supplemented with 1% non-essential amino acids and containing 10% foetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, 100mg/mL streptomycin, 1 mM Hepes
Media Volume10 mL
AssayCell viability (lactate dehydrogenase assay), membrane integrity (TEER), tight junction formation (immunostaining for ZO-1), fluorescein permeability
Pagliari et al (2014) - Generating a vascularised cardiac tissue construct with cardiomyocyte-like and endothelial-like cells, using a 3D scaffold and dynamic culture
Cell Types
Human mesenchymal stem cells (hMSC), human cardiomyocyte progenitor cells (hCMPC)
Cell SourceLonza; isolated from healthy donor atrial biopsy
Flow Rate (µL/min)200
Shear Pressure (Pa)1.1 x 10−^6
Time in flow7 days
Culture config/substratePorous gelatin scaffold soaked in Matrigel
Seeding Density2x10^5 cells per cover slip
MediaCardiac differentiation medium: IMDM/Ham’s-F12 1:1 with L-glutamine, 2% horse serum, non-essential amino acids, insulin-transferrin-selenium, penicillin/streptomycin
Media Volume21 mL
AssayqRT-PCR, FACS analysis, fluorescence microscopy
Mattei, Giusti & Ahluwalia (2014) - Physiologically relevant in vitro bioreactor design: using computational fluid dynamics to compare milli- and micro-fluidic systems
Cell Types
Hepatocytes
Cell Source
Flow Rate (µL/min)180
Shear Pressure (Pa)5.2 x 10-^6
Time in flow
Culture config/substrate0.2mm thick hydrogel construct
Seeding Density2.47x10^6 cells per chamber
Media
Media Volume
AssayComputational fluid dynamic modelling
Ucciferri et al (2014) - Toxicity of nanoparticles and the role of flow in endothelial cell response in vitro

Cell Types
Primary human endothelial cells (HUVECs)
Cell SourceIsolated from umbilical vein
Flow Rate (µL/min)100
Shear Pressure (Pa)1x10−^6
Time in flow24 hours
Culture config/substrate1% gelatin-coated 13mm-diameter plastic slide
Seeding Density2x10^4 cells/cm2
MediaEagle's minimal essential medium with supplements
Media Volume15 mL
AssayAssays for markers used in nanotoxicty screening
Saha & Miranda-Azpiazu et al (2015)
Blood brain barrier model
Cell Types
Human brain primary astrocytes, human brain primary vascular pericytes, human brain primary endothelial cells
Cell SourceIsolated from human brain
Flow Rate (µL/min)40
Shear Pressure (Pa)
Time in flow
Culture config/substrateGlass coated coverslip (astrocytes), Transwell membrane (pericytes and endothelial cells)
Seeding Density
MediaDifferent or conditioned media
Media Volume
AssayMTT assay (viability)
Ramachandran et al (2015) - Generation and long-term culture of 3D liver organoids using upcytes

Cell Types
Human upcyte hepatocytes, human upcyte liver sinusoildal endothelial cells, human upcyte mesenchymal stem cells
Cell SourceMedicyte
Flow Rate (µL/min)
Shear Pressure (Pa)
Time in flow10 days

Culture config/substrateSuspended in media then added to Matrigel-coated chamber
Seeding Density1x10^6 hepatocytes, 1x10^6 LSECs and 0.2x10^6 MSCs mixed in 1 mL liver organoid growth medium
MediaLiver organoid growth medium: upctye Hepatocyte Growth Medium and upcyte LSEC Growth Medium in 1:1 ratio
Media Volume
AssayIn situ hybridisation, CYP activity assay, immunocytochemistry, immunohistochemistry, qRT-PCR
Faure et al (2016) - Effect of metformin on chicken Sertoli and germ cells

Cell Types
Seminiferous tubule culture containing germ cells preserved on polarised Sertoli cells
Cell Source6-week-old male chickens
Flow Rate (µL/min)250
Shear Pressure (Pa)
Time in flow96 hours
Culture config/substratePolyethylene terephthalate insert
Seeding Density1.2x10^5 cells per insert
MediaF12/DMEM supplemented with antibiotics, 500 µM pyruvate, 10 µg/mL transferrin, 107 M testosterone, 17.6 µg/mL vitamin C, 10 µg/mL vitamin E, 86 ng/mL vitamin A, 5 ng/mL, oFSH, 1.109 recombinant human insulin-like growth factor 1, 1% FCS
Media Volume
AssayImmunolabelling for flow cytometric analysis
Rashidi et al (2016) - Using fluid shear stress to improve hepatocyte-like cell function
Cell Types
Human embryonic stem cell-derived hepatocyte-like cells
Cell SourceHuman embryo
Flow Rate (µL/min)100 & 200
Shear Pressure (Pa)2.9 x 10-^6 & 4.7 x 10-^6
Time in flow18 hours
Culture config/substrateMatrigel-coated Thermanox coverslip
Seeding Density
Media
Media Volume
AssayCytochrome P450 assay (Cyp1A2 activity), viability (ATP levels measured, indicative of Cyp2D6 activity), albumin & alpha-fetoprotein ELISA
Shannahan et al (2016) - Influence of iron oxide biocorona (layer of biomolecules adsorbed to surface of nanoparticle) on endothelial cells under physiological conditions
Cell Types
Rat aortic endothelial cells
Cell SourceCell Applications Inc.
Flow Rate (µL/min)1000
Shear Pressure (Pa)1 x 10-^5
Time in flow1-2 hours
Culture config/substrateGlass coverslip
Seeding Density
MediaRat endothelial cell growth medium
Media Volume
AssayInductively coupled plasma mass spectrometry, darkfield microscopy, PCR array, qRT-PCR, fluorescent microscopy
Pedersen et al (2016) - Evaluation of flow systems for hepatocyte culture

Cell Types
Primary rat hepatocytes
Cell SourceFreshly isolated
Flow Rate (µL/min)500
Shear Pressure (Pa)4 x 10-^6 - 5.7 x 10-^5
Time in flow31 days

Culture config/substrateAlginate bead (250, 500 or 1000 µm diameter, 50% v/v hepatocytes)
Seeding Density1.5 million (total number of cells per chamber)
MediaWilliams' E medium
Media Volume
AssayViability (acridine orange/propidium iodide staining)
Nithiananthan et al (2016) - Fibroblast differentiation in response to flow


Cell Types
Normal oral fibroblasts; normal human dermal fibroblasts
Cell SourceWaste tissues from oral surgical procedures; Promocell
Flow Rate (µL/min)75 & 150
Shear Pressure (Pa)3.61 x 10-^6 (at 150 µL/min)
Time in flow24 hours
Culture config/substrateGlass coverslip

Seeding Density
MediaDMEM containing 0.5% FCS

Media Volume
AssayMicroarray analysis, immunofluorescence staining, qRT-PCR, immunoblotting, ELISA

Martin et al (2017) - Challenging the hypothesis that branching morphogenesis in kidney epithelia is caused by a secreted autocrine inhibitor of cell motility

Cell Types
Canine renal cells (MDCK II)

Cell SourceEuropean Collection of Cell Cultures
Flow Rate (µL/min)3000
Shear Pressure (Pa)5 x 10-^5 - 5 x 10-^4
Time in flow1.5 hours
Culture config/substrateMicro-patterned 1 cm2 glass plate consisting of amino-terminated adhesion-permissive substrate surrounded by adhesion-resistant substrate

Seeding Density9x10^5 cells in suspension per glass plate
MediaEagle's minimal essential medium supplemented with 2mM L-glutamine, 100 U penicillin/streptomycin, 5% foetal calf serum
Media Volume
AssayLive imaging, image analysis
Lewis et al - Use of flow & 3D scaffolds in cell culture and hepatocyte response to cyclophosphamide

Cell Types
NCI-H292 human lung carcinoma cells, HepG2 hepatocellular carcinoma cells, rat primary hepatocytes
Cell Source
Flow Rate (µL/min)
Shear Pressure (Pa)
Time in flow24 hours; 3 days (HepG2)
Culture config/substrateGlass coverslip (coated with 0.1 mg/mL collagen) or Alvetex scaffold, then coated with 1 mg/mL collagen
Seeding Density0.2x10^6; 0.3x10^6
MediaRPMI medium
Media Volume
AssayViability (alamarBlue resazurin reduction assay, MTT assay); glucose, lactic acid and albumin levels; light microscopy
Da-silva et al - Using Quasi Vivo (2D fluidic monoculture) to assess compounds that have low intrinsic clearance


Cell Types
Cryopreserved primary human hepatocytes


Cell SourceIsolated from donors
Flow Rate (µL/min)150
Shear Pressure (Pa)3.8 x 10-^6
Time in flow7 days
Culture config/substrateCollagen coated

Seeding Density1.5x10^5 cells/coverslip

MediaWilliams' E medium & supplements (ITS, Glutamax, Dexamethasone, Gentamicine)
Media Volume
AssayElimination rate of test compounds, qRT-PCR
Yoon et al - Non-specific binding in Quasi Vivo system with different types of tubing
Cell Types
Cell Source
Flow Rate (µL/min)
Shear Pressure (Pa)
Time in flow4 hours, 2.5 hours
Culture config/substrate
Seeding Density
MediaMedia containing 10 µM 7-ethoxy-coumarin
Media Volume20 mL
AssayAdherence of 7EC, release of 7EC, 3D CFD modelling to predict distribution of small molecules

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