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Endogro mv complete media

Manufactured by Merck Group
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Sourced in United Kingdom

EndoGRO-MV Complete Media is a pre-formulated cell culture medium designed to support the growth and maintenance of human microvascular endothelial cells. The medium contains essential nutrients, growth factors, and other components required for the in vitro culture of these cells.

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Lab products found in correlation

H 6023, by Cell Biologics (2 mentions) Scc0066, by Merck Group (2 mentions) C57 6023, by Cell Biologics (2 mentions) Enspire plate reader, by PerkinElmer (2 mentions) Leibovitz l 15 media, by Lonza (1 mentions) Collagen type 1, by Merck Group (1 mentions) Evom2 voltohmmeter, by World Precision Instruments (1 mentions) Penicillin, by Merck Group (1 mentions) Streptomycin, by Merck Group (1 mentions) Hcmec d3 cells, by Merck Group (1 mentions)

Close spelling variants of this product

EndoGRO (16 citations) EndoGRO™-MV Complete Media Kit (12 citations) EndoGRO-MV Complete Culture Media Kit (9 citations) EndoGRO-MV-VEGF Complete Culture Media Kit (5 citations) EndoGROTM-MV-VEGF Complete Media Kit (2 citations) EndoGRO-MV Complete Culture Media (2 citations) EndoGRO™-MV-VEGF Complete Media Kit medium (1 citations)

6 protocols using endogro mv complete media

1

Culturing Human Cerebral Microvascular Endothelial Cells

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The human cerebral microvascular endothelial cell (hCMEC) line (Merck, London, UK) was cultured in EndoGRO-MV Complete Media (Merck) supplemented with 1% penicillin–streptomycin and grown at 37°C with 5% CO2 in culture vessels coated with 5 µg/cm2 Collagen Type 1 (Merck). Prior to the experiment, the cells were seeded onto coverslips in a 6-well plate or a fluorodisc in culture media at a density of 4 × 104 cells/ml and left for 24 h to settle. The culture media was then replaced with Leibovitz L-15 media (Lonza), and the cells were moved to a 37°C incubator suitable for bacterial infection.
Dhanoa G.K., Kushnir I., Qimron U., Roper D.I, & Sagona A.P. (2022). Investigating the effect of bacteriophages on bacterial FtsZ localisation. Frontiers in Cellular and Infection Microbiology, 12, 863712.
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2

Transcellular Permeability Assay of RS194B

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Traversal of RS194B across brain endothelial cells was assessed using Human cerebral microvascular cells (HCMEC/D3), obtained from Cornell University. Cells were grown in EndoGRO-MV Complete media (EMD Millipore). For transcellular assays, cells were seeded at approximately 5 × 104 cells/cm2 in 12-well transwell tissue culture inserts (0.4 μm pore size, polycarbonate membrane) (Corning Life Sciences). All inserts were collagen coated prior to use. Cells were seeded in media containing 10 mM LiCl, and media was exchanged every 2-3 days. Monolayer integrity was monitored continuously via transendothelial electrical resistance using the EVOM2 voltohmmeter (World Precision Instruments). Inserts were used in assays 6-7 days following seeding. Compound was dissolved at 100 μM in HBSS for testing and added to the upper (apical) well of the insert. Buffer samples were removed from the lower (basolateral) well at 0, 15, 30, 60, 90, and 120 minutes after dosing. Each compound was examined in at least three replicates. As above, compound in buffer was quantified by UPLC as previously described (Bennion et al, 2017 ). Permeability of each compound across inserts not containing cells was examined as a control in each assay plate. Effective permeability (Peff [cm/s]) was calculated using the permeability surface area product method, as described by Deli et al.
Malfatti M.A., Enright H.A., Be N.A., Kuhn E.A., Hok S., McNerney M.W., Lao V., Nguyen T.H., Lightstone F.C., Carpenter T.S., Bennion B.J, & Valdez C.A. (2017). The biodistribution and pharmacokinetics of the oxime acetylcholinesterase reactivator RS194B in guinea pigs. Chemico-biological interactions, 277, 159-167.
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3

Cultivation of hCMEC/D3 Blood-Brain Barrier Cells

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The blood-brain barrier hCMEC/D359 (link) (human cerebral microvascular endothelial cells) cell line (Merck, UK) consists of enriched cerebral microvascular endothelial cells immortalised by lentiviral vector transduction with the catalytic subunit of human telomerase (hTERT) and SV40 large T antigen.
hCMECs were cultured in EndoGRO-MV Complete Media (Merck) supplemented with 1 ng/ml bFGF (FGF-2) (Merck), 100 IU/ml Penicillin (Sigma-Aldrich), and 100 μg/ml Streptomycin (Sigma-Aldrich) and maintained under a humidified atmosphere at 37 C in 5% CO2. All culture vessels were coated with 5 μg/cm2 Collagen Type 1 (Collagen-1) (Merck) in PBS for 1 h at 37 C before use.
Møller-Olsen C., Ross T., Leppard K.N., Foisor V., Smith C., Grammatopoulos D.K, & Sagona A.P. (2020). Bacteriophage K1F targets Escherichia coli K1 in cerebral endothelial cells and influences the barrier function. Scientific Reports, 10, 8903.
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4

AAV transduction efficiency in brain endothelial cells

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Multiple lots of mBMVEC and hBMVEC cells were obtained from CellBiologics (H-6023 & C57-6023) and maintained in endothelial cell media (H1168 & M1168). hCMEC/D3 cells were obtained from Millipore (SCC0066) and maintained in EndoGRO™-MV Complete Media (SCME004). All cells were handled according to the manufacturer’s instructions. For the Luciferase assays, 5000 cells/well were seeded in 96 well plates (PerkinElmer, 6005680). One day later, AAV9, AAV-PHP.eB or AAV-BI30 carrying pAAV-CAG-eGFP-p2A-luciferase was added at 20,000 vg/cell. 24 hours after transduction, a luciferase reporter assay was performed according to the manufacturer’s instructions (PerkinElmer, 6066761) on an EnSpire plate reader (PerkinElmer). For flow cytometry, hCMEC/D3 cells were plated at 434,000 cells/well in 24 well plates and exposed to the indicated dose of AAV-BI30 or AAV9. The media was exchanged for fresh media after 24 hours and transduction was assessed at 4 days post-administration on a Beckman CytoFLEX S Flow Cytometer using FlowJo 10.8.1 (BD Biosciences).
Krolak T., Chan K.Y., Kaplan L., Huang Q., Wu J., Zheng Q., Kozareva V., Beddow T., Tobey I.G., Pacouret S., Chen A.T., Chan Y.A., Ryvkin D., Gu C, & Deverman B.E. (2022). A High-Efficiency AAV for Endothelial Cell Transduction Throughout the Central Nervous System. Nature cardiovascular research, 1(4), 389-400.
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5

AAV transduction efficiency in brain endothelial cells

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Multiple lots of mBMVEC and hBMVEC cells were obtained from CellBiologics (H-6023 & C57-6023) and maintained in endothelial cell media (H1168 & M1168). hCMEC/D3 cells were obtained from Millipore (SCC0066) and maintained in EndoGRO™-MV Complete Media (SCME004). All cells were handled according to the manufacturer’s instructions. For the Luciferase assays, 5000 cells/well were seeded in 96 well plates (PerkinElmer, 6005680). One day later, AAV9, AAV-PHP.eB or AAV-BI30 carrying pAAV-CAG-eGFP-p2A-luciferase was added at 20,000 vg/cell. 24 hours after transduction, a luciferase reporter assay was performed according to the manufacturer’s instructions (PerkinElmer, 6066761) on an EnSpire plate reader (PerkinElmer). For flow cytometry, hCMEC/D3 cells were plated at 434,000 cells/well in 24 well plates and exposed to the indicated dose of AAV-BI30 or AAV9. The media was exchanged for fresh media after 24 hours and transduction was assessed at 4 days post-administration on a Beckman CytoFLEX S Flow Cytometer using FlowJo 10.8.1 (BD Biosciences).
Krolak T., Chan K.Y., Kaplan L., Huang Q., Wu J., Zheng Q., Kozareva V., Beddow T., Tobey I.G., Pacouret S., Chen A.T., Chan Y.A., Ryvkin D., Gu C, & Deverman B.E. (2022). A High-Efficiency AAV for Endothelial Cell Transduction Throughout the Central Nervous System. Nature cardiovascular research, 1(4), 389-400.
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6

Bacterial adhesion and invasion assay

Cited 3 times
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hCMEC/D3 cells (obtained from Millipore) were grown in EndoGRO-MV complete media (Millipore, SCME004) supplemented with 5% fetal bovine serum and fibroblast growth factor 2 (1 ng/ml; Millipore). Cells were grown in tissue culture treated 24-well plates and 5% CO2 at 37°C. Assays to determine the total number of bacteria adhered to host cells or intracellular bacteria were performed using GBS empty vector and complement strain as described previously (24 (link), 31 (link), 32 (link)). See Supplementary Text for more details.
Joyce L.R., Kim S., Spencer B.L., Christensen P.M., Palmer K.L., Guan Z., Siegenthaler J.A, & Doran K.S. (2024). Streptococcus agalactiae glycolipids promote virulence by thwarting immune cell clearance. Science Advances, 10(22), eadn7848.
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