QUASI VIVO® GLOSSARY
Check out a compilation of information taken from publications and posters featuring Quasi Vivo®.
| Martin et al (2017) - Challenging the hypothesis that branching morphogenesis in kidney epithelia is caused by a secreted autocrine inhibitor of cell motility |
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|---|---|---|
| Cell Types | Canine renal cells (MDCK II) |
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| Cell Source | European Collection of Cell Cultures |
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| Flow Rate (µL/min) | 3000 |
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| Shear Pressure (Pa) | 5 x 10-^5 - 5 x 10-^4 |
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| Time in flow | 1.5 hours |
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| Culture config/substrate | Micro-patterned 1 cm2 glass plate consisting of amino-terminated adhesion-permissive substrate surrounded by adhesion-resistant substrate |
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| Seeding Density | 9x10^5 cells in suspension per glass plate |
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| Media | Eagle's minimal essential medium supplemented with 2mM L-glutamine, 100 U penicillin/streptomycin, 5% foetal calf serum | |
| Media Volume | ||
| Assay | Live imaging, image analysis |
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| Nithiananthan et al (2016) - Fibroblast differentiation in response to flow |
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|---|---|---|
| Cell Types | Normal oral fibroblasts; normal human dermal fibroblasts |
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| Cell Source | Waste tissues from oral surgical procedures; Promocell |
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| Flow Rate (µL/min) | 75 & 150 |
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| Shear Pressure (Pa) | 3.61 x 10-^6 (at 150 µL/min) |
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| Time in flow | 24 hours |
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| Culture config/substrate | Glass coverslip |
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| Seeding Density | ||
| Media | DMEM containing 0.5% FCS |
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| Media Volume | ||
| Assay | Microarray analysis, immunofluorescence staining, qRT-PCR, immunoblotting, ELISA |
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| Pedersen et al (2016) - Evaluation of flow systems for hepatocyte culture |
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|---|---|---|
| Cell Types | Primary rat hepatocytes |
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| Cell Source | Freshly isolated |
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| Flow Rate (µL/min) | 500 |
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| Shear Pressure (Pa) | 4 x 10-^6 - 5.7 x 10-^5 |
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| Time in flow | 31 days |
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| Culture config/substrate | Alginate bead (250, 500 or 1000 µm diameter, 50% v/v hepatocytes) |
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| Seeding Density | 1.5 million (total number of cells per chamber) |
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| Media | Williams' E medium |
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| Media Volume | ||
| Assay | Viability (acridine orange/propidium iodide staining) |
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| Shannahan et al (2016) - Influence of iron oxide biocorona (layer of biomolecules adsorbed to surface of nanoparticle) on endothelial cells under physiological conditions |
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|---|---|---|
| Cell Types | Rat aortic endothelial cells |
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| Cell Source | Cell Applications Inc. |
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| Flow Rate (µL/min) | 1000 |
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| Shear Pressure (Pa) | 1 x 10-^5 |
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| Time in flow | 1-2 hours |
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| Culture config/substrate | Glass coverslip |
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| Seeding Density | ||
| Media | Rat endothelial cell growth medium |
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| Media Volume | ||
| Assay | Inductively coupled plasma mass spectrometry, darkfield microscopy, PCR array, qRT-PCR, fluorescent microscopy |
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| Rashidi et al (2016) - Using fluid shear stress to improve hepatocyte-like cell function |
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|---|---|---|
| Cell Types | Human embryonic stem cell-derived hepatocyte-like cells |
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| Cell Source | Human embryo |
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| Flow Rate (µL/min) | 100 & 200 |
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| Shear Pressure (Pa) | 2.9 x 10-^6 & 4.7 x 10-^6 |
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| Time in flow | 18 hours |
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| Culture config/substrate | Matrigel-coated Thermanox coverslip |
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| Seeding Density | ||
| Media | ||
| Media Volume | ||
| Assay | Cytochrome P450 assay (Cyp1A2 activity), viability (ATP levels measured, indicative of Cyp2D6 activity), albumin & alpha-fetoprotein ELISA |
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| Faure et al (2016) - Effect of metformin on chicken Sertoli and germ cells |
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|---|---|---|
| Cell Types | Seminiferous tubule culture containing germ cells preserved on polarised Sertoli cells |
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| Cell Source | 6-week-old male chickens |
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| Flow Rate (µL/min) | 250 |
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| Shear Pressure (Pa) | ||
| Time in flow | 96 hours |
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| Culture config/substrate | Polyethylene terephthalate insert |
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| Seeding Density | 1.2x10^5 cells per insert |
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| Media | F12/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 |
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| Media Volume | ||
| Assay | Immunolabelling for flow cytometric analysis |
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| Ramachandran et al (2015) - Generation and long-term culture of 3D liver organoids using upcytes |
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|---|---|---|
| Cell Types | Human upcyte hepatocytes, human upcyte liver sinusoildal endothelial cells, human upcyte mesenchymal stem cells |
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| Cell Source | Medicyte |
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| Flow Rate (µL/min) | ||
| Shear Pressure (Pa) | ||
| Time in flow | 10 days |
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| Culture config/substrate | Suspended in media then added to Matrigel-coated chamber | |
| Seeding Density | 1x10^6 hepatocytes, 1x10^6 LSECs and 0.2x10^6 MSCs mixed in 1 mL liver organoid growth medium | |
| Media | Liver organoid growth medium: upctye Hepatocyte Growth Medium and upcyte LSEC Growth Medium in 1:1 ratio |
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| Media Volume | ||
| Assay | In situ hybridisation, CYP activity assay, immunocytochemistry, immunohistochemistry, qRT-PCR |
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| Saha & Miranda-Azpiazu et al (2015) Blood brain barrier model |
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|---|---|---|
| Cell Types | Human brain primary astrocytes, human brain primary vascular pericytes, human brain primary endothelial cells |
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| Cell Source | Isolated from human brain |
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| Flow Rate (µL/min) | 40 |
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| Shear Pressure (Pa) | ||
| Time in flow | ||
| Culture config/substrate | Glass coated coverslip (astrocytes), Transwell membrane (pericytes and endothelial cells) | |
| Seeding Density | ||
| Media | Different or conditioned media |
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| Media Volume | ||
| Assay | MTT assay (viability) |
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| Ucciferri et al (2014) - Toxicity of nanoparticles and the role of flow in endothelial cell response in vitro |
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|---|---|---|
| Cell Types | Primary human endothelial cells (HUVECs) | |
| Cell Source | Isolated from umbilical vein |
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| Flow Rate (µL/min) | 100 | |
| Shear Pressure (Pa) | 1x10−^6 | |
| Time in flow | 24 hours |
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| Culture config/substrate | 1% gelatin-coated 13mm-diameter plastic slide |
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| Seeding Density | 2x10^4 cells/cm2 |
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| Media | Eagle's minimal essential medium with supplements |
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| Media Volume | 15 mL |
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| Assay | Assays for markers used in nanotoxicty screening |
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| Mattei, Giusti & Ahluwalia (2014) - Physiologically relevant in vitro bioreactor design: using computational fluid dynamics to compare milli- and micro-fluidic systems |
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|---|---|---|
| Cell Types | Hepatocytes |
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| Cell Source | ||
| Flow Rate (µL/min) | 180 |
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| Shear Pressure (Pa) | 5.2 x 10-^6 |
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| Time in flow | ||
| Culture config/substrate | 0.2mm thick hydrogel construct |
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| Seeding Density | 2.47x10^6 cells per chamber |
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| Media | ||
| Media Volume | ||
| Assay | Computational fluid dynamic modelling |
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| 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 Source | Lonza; isolated from healthy donor atrial biopsy | |
| Flow Rate (µL/min) | 200 | |
| Shear Pressure (Pa) | 1.1 x 10−^6 | |
| Time in flow | 7 days | |
| Culture config/substrate | Porous gelatin scaffold soaked in Matrigel | |
| Seeding Density | 2x10^5 cells per cover slip | |
| Media | Cardiac differentiation medium: IMDM/Ham’s-F12 1:1 with L-glutamine, 2% horse serum, non-essential amino acids, insulin-transferrin-selenium, penicillin/streptomycin |
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| Media Volume | 21 mL |
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| Assay | qRT-PCR, FACS analysis, fluorescence microscopy | |
| Giusti et al (2014) - Dual-flow intestinal barrier model | ||
|---|---|---|
| Cell Types | Caco-2/TC7 intestinal epithelial cells | |
| Cell Source | Sigma-Aldrich |
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| Flow Rate (µL/min) | Apical = 200, basal = 100 |
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| Shear Pressure (Pa) | 6 x 10-^4 | |
| Time in flow | 24 & 48 hours | |
| Culture config/substrate | Monolayer on polycarbonate membrane of QV600 insert |
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| Seeding Density | 3x10^4 cells/cm2 | |
| Media | Dulbecco’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 Volume | 10 mL |
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| Assay | Cell viability (lactate dehydrogenase assay), membrane integrity (TEER), tight junction formation (immunostaining for ZO-1), fluorescein permeability |
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| 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/substrate | Ultrathin silicon nitride membrane with built-in TEER electrodes (for intestinal epithelial cell culture) | |
| Seeding Density | ||
| Media | EMEM-GF (common medium for all 3 cell types) | |
| Media Volume | ||
| Assay | Phenotypic 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) | |
| 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 Source | Laboratory 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 flow | 15, 24 & 48 hours | |
| Culture config/substrate | Collagen-coated PLGA scaffold, coated with alginate film; nylon mesh | |
| Seeding Density | 1x10^5 hepatocytes per scaffold | |
| Media | Endothelial cell growth medium containing 10% FBS | |
| Media Volume | Endothelial cell growth medium containing 10% FBS | |
| Assay | Viability, morphology; glycerol, D-lactate, L-alanine (spectrophotometry); free fatty acids, glucose (enzyme assays); IL-6, albumin, E-selectin (ELISA) | |
| 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 |
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| Shear Pressure (Pa) | 2.9 x 10-^6 | |
| Time in flow | 24 hours | |
| Culture config/substrate | Coated in collagen gel |
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| Seeding Density | 2x10^5 cells per well | |
| Media | Serum-free Williams' medium | |
| Media Volume | ||
| Assay | Almar blue vitality | |
| 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 Source | Laboratory 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 flow | 15, 24 & 48 hours | |
| Culture config/substrate | Collagen-coated PLGA scaffold, coated with alginate film; nylon mesh | |
| Seeding Density | 1x10^5 hepatocytes per scaffold | |
| Media | Conditioned endothelial cell growth medium containing 10% FBS | |
| Media Volume | 15 mL |
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| Assay | Glucose, free fatty acid, glycerol, D-lactate, L-alanine and albumin concentration |
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| Vozzi et al (2011) - In vitro tissue and organ modelling using connected bioreactors |
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|---|---|---|
| Cell Types | HepG2 hepatocytes | |
| Cell Source | Human hepatoblastoma line | |
| Flow Rate (µL/min) | 250 | |
| Shear Pressure (Pa) | 5 x 10-^6 | |
| Time in flow | 1 week; 48 hours | |
| Culture config/substrate | Collagen-coated PLGA scaffold, coated with alginate film | |
| Seeding Density | 1x10^5 cells per scaffold | |
| Media | Conditioned endothelial cell growth medium containing 10% FBS | |
| Media Volume | 15 mL |
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| Assay | Glucose, free fatty acid, glycerol, D-lactate, L-alanine and albumin concentration | |
| Vinci et al (A) (2011) - Liver detoxification gene expression and activity under flow | ||
|---|---|---|
| Cell Types | Primary human hepatocytes |
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| Cell Source | Isolated from liver donors |
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| Flow Rate (µL/min) | 250 - 500 |
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| Shear Pressure (Pa) | 5 x 10-^6 - 1 x 10-^5 |
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| Time in flow | 7 - 21 days |
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| Culture config/substrate | Collagen sandwich |
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| Seeding Density | ~1.9x10^5 cells per cover slip |
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| Media | Long 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 | ||
| Assay | qRT-PCR, drug metabolism, albumin & urea secretion |
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| 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 Source | Adult 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 flow | 24 hours | |
| Culture config/substrate | Collagen sandwich | |
| Seeding Density | 2x10^5 cells per cover slip | |
| Media | FBS-free complete Williams' E | |
| Media Volume | 10 mL | |
| Assay | Viability, albumin ELISA | |
| Lewis et al - Use of flow & 3D scaffolds in cell culture and hepatocyte response to cyclophosphamide |
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|---|---|---|
| Cell Types | NCI-H292 human lung carcinoma cells, HepG2 hepatocellular carcinoma cells, rat primary hepatocytes |
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| Cell Source | ||
| Flow Rate (µL/min) | ||
| Shear Pressure (Pa) | ||
| Time in flow | 24 hours; 3 days (HepG2) |
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| Culture config/substrate | Glass coverslip (coated with 0.1 mg/mL collagen) or Alvetex scaffold, then coated with 1 mg/mL collagen |
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| Seeding Density | 0.2x10^6; 0.3x10^6 |
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| Media | RPMI medium |
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| Media Volume | ||
| Assay | Viability (alamarBlue resazurin reduction assay, MTT assay); glucose, lactic acid and albumin levels; light microscopy |
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| Da-silva et al - Using Quasi Vivo (2D fluidic monoculture) to assess compounds that have low intrinsic clearance |
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|---|---|---|
| Cell Types | Cryopreserved primary human hepatocytes |
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| Cell Source | Isolated from donors |
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| Flow Rate (µL/min) | 150 |
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| Shear Pressure (Pa) | 3.8 x 10-^6 |
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| Time in flow | 7 days |
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| Culture config/substrate | Collagen coated |
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| Seeding Density | 1.5x10^5 cells/coverslip |
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| Media | Williams' E medium & supplements (ITS, Glutamax, Dexamethasone, Gentamicine) |
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| Media Volume | ||
| Assay | Elimination rate of test compounds, qRT-PCR |
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| 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 flow | 4 hours, 2.5 hours |
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| Culture config/substrate | ||
| Seeding Density | ||
| Media | Media containing 10 µM 7-ethoxy-coumarin |
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| Media Volume | 20 mL |
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| Assay | Adherence of 7EC, release of 7EC, 3D CFD modelling to predict distribution of small molecules |
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