Human umbilical vein endothelial cells (HUVECs, Lonza) and red fluorescent protein-expressing HUVECs (RFP-HUVECs) (Angio-Proteomie) were cultured in endothelial basal medium-2 (EBM-2, Lonza) supplemented with EGM-2 bullet kit and normal human lung fibroblasts (LFs, Lonza) were cultured in fibroblast basal medium (FBM-2, Lonza) supplemented with FGM-2 bullet kit. All cancer cells were from Korean cell line bank (KCLB, Korea). Human colon cancer (SW620) and human gastric cancer (MKN74) were cultured in Dulbescco's modified Eagle's mideiun (DMEM, Hyclone) supplemented with 10% fetal bovine serum (FBS, Gibco), penicillin, and streptomycin (100 U/mL) (Sigma Aldrich). All cells were incubated at 37°C in a humidified 5% CO2 atmosphere.
Fbm 2
Manufactured by Lonza
The FBM-2 is a laboratory equipment designed for cell culture applications. It serves as a Fluidized Bed Mixer, providing a controlled and homogeneous mixing environment for cell suspension cultures.
Automatically generated - may contain errors
Lab products found in correlation
Rfp huvecs, by Angio-Proteomie (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Penicillin, by Merck Group (2 mentions)
Streptomycin, by Merck Group (2 mentions)
Ebm 2, by Lonza (2 mentions)
Huvecs, by Lonza (2 mentions)
Fibronectin, by Merck Group (1 mentions)
Fgm 2, by Lonza (1 mentions)
Normal human lung fibroblasts nhlfs, by Lonza (1 mentions)
Pcs 100 010, by American Type Culture Collection (1 mentions)
Cc 3132, by Lonza (1 mentions)
Cc 2509, by Lonza (1 mentions)
Cc 3202, by Lonza (1 mentions)
Egm 2mv, by Lonza (1 mentions)
4 protocols using fbm 2
1
In Vitro Cell Culture Protocol
2
2D Culture of Cancer Cells with Fibronectin
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Human umbilical vein endothelial cells (HUVECs, Lonza) and red fluorescent protein-expressing HUVECs (RFP-HUVECs) (Angio-Proteomie) were cultured in endothelial basal medium-2 (EBM-2, Lonza) supplemented with EGM-2 bullet kit and normal human lung fibroblasts (LFs, Lonza) were cultured in fibroblast basal medium (FBM-2, Lonza) supplemented with FGM-2 bullet kit. All cancer cells were from Korean cell line bank (KCLB, Korea). Human colon cancer (SW620) and human gastric cancer (MKN74) were cultured in Dulbescco's modified Eagle's mideiun (DMEM, Hyclone) supplemented with 10% fetal bovine serum (FBS, Gibco), penicillin, and streptomycin (100 U/mL) (Sigma Aldrich). All cells were incubated at 37°C in a humidified 5% CO2 atmosphere.
For 2D assay, PDMS block was punched using 6 mm biopsy punch and pre-treated with 10 μg/mL of fibronectin (Sigma) for 2 h and washed 3 times before cancer cell seeding. Cancer cells were seeded with 100 μL of the 1.5 × 104 cells/ml density into each well. After 2 days of DMEM media culture, culture media was changed into different contained media with different concentration (0.0%, 0.2%, and 0.4%) and analyzed after 3 days of culture.
For 2D assay, PDMS block was punched using 6 mm biopsy punch and pre-treated with 10 μg/mL of fibronectin (Sigma) for 2 h and washed 3 times before cancer cell seeding. Cancer cells were seeded with 100 μL of the 1.5 × 104 cells/ml density into each well. After 2 days of DMEM media culture, culture media was changed into different contained media with different concentration (0.0%, 0.2%, and 0.4%) and analyzed after 3 days of culture.
3
Fibroblast Maintenance and Serum Starvation
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Normal human lung fibroblasts (NHLFs) were purchased from Lonza (Allendale, NJ) and maintained in fibroblast growth medium 2 (FGM-2; Lonza) for experiments until passage 6 per the provider’s instruction. Prior to treatment, cells that had reached 80% confluence were serum starved in fibroblast basal medium 2 (FBM-2; Lonza) with 0.2% bovine serum albumin (BSA) overnight [17 (link)].
4
HUVECs and NHDFs for Talin Abundance Analysis
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HUVECs (PCS-100-010, ATCC) and NHDFs (CC-2509, Lonza) as a control for talin abundance were purchased and maintained in endothelial cell growth medium (EGM-2MV; CC-3202) or fibroblast growth medium (FBM-2; CC-3132, both from Lonza). Both culture media contained 100 U/mL penicillin/streptomycin (P/S; #15140, Gibco, Grand Island, NY, USA). All cells were incubated at 37 °C in a humidified incubator with 5% CO2 and used at passage 7 to 10. Culture media were changed every two days. In experiments using KCH-1521, HUVECs were incubated with either KCH-1521 at a concentration of 100 μM or the equivalent amount of DMSO (1% (v/v)) as a vehicle control in culture medium. The cell length in the long axis and cell numbers were measured by phase-contrast images using the ImageJ (ver. 1.49) program (National Institute of Health, Bethesda, MD, USA).
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