Primary human lung microvascular endothelial cells (HMVEC-Ls) and human umbilical vein endothelial cells (HUVECs) were obtained from Lonza (Basel, Switzerland) and cultured in endothelial growth medium-2 supplemented with human epidermal growth factor (hEGF), human vascular endothelial growth factor (hVEGF), human fibroblast growth factor basic (hEGFb), an analog of human insulin-like growth factor-1 (R3-IGF-1), ascorbic acid, hydrocortisone, fetal bovine serum (FBS), heparin and gentamicin/amphotericin-B (GA) (Lonza). Mouse VECs line UV♀2 cells were obtained from the RIKEN BioResource Research Center (RCB1994, Tukuba, Japan), and cultured in Dulbecco's modified eagle medium (DMEM; Fujifilm Wako, Osaka, Japan) with 10% FBS (Thermo Fisher Scientific, Waltham, MA, USA), 1 mM glutamine, 100 U/mL penicillin and 100 µg/mL streptomycin (all from Fujifilm Wako). Cells were cultured in a 5% CO 2 incubator at 37 • C, and cells at passages 3-8 were used for all experiments.
Gentamicin amphotericin b ga
Manufactured by Lonza
Sourced in Switzerland, United States
Gentamicin/amphotericin-B (GA) is a laboratory product that combines two antimicrobial agents, gentamicin and amphotericin-B. It is used as a supplement in cell culture media to prevent bacterial and fungal contamination.
Automatically generated - may contain errors
Lab products found in correlation
Heparin, by Lonza (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Lonza (2 mentions)
Glutamine, by Fujifilm (1 mentions)
Penicillin, by Fujifilm (1 mentions)
Streptomycin, by Fujifilm (1 mentions)
Dulbecco s modified eagle medium dmem, by Fujifilm (1 mentions)
Hmvec l, by Lonza (1 mentions)
Huvecs, by Lonza (1 mentions)
Dmem aq, by Merck Group (1 mentions)
Hfgf b, by Lonza (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Hydrocortisone, by Lonza (1 mentions)
Ascorbic acid, by Lonza (1 mentions)
Trypsin edta, by Merck Group (1 mentions)
Dexamethasone (dex), by Lonza (1 mentions)
Skeletal muscle cell growth medium 2, by Lonza (1 mentions)
4 protocols using gentamicin amphotericin b ga
1
Endothelial Cell Culture Protocol
2
Culturing BAE and HAEC Cells
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BAE cells were cultured in Dulbecco's Modified Eagle's Medium-AQ (DMEM-AQ, Sigma Aldrich) supplemented with 10% heat inactivated fetal bovine serum (FBS; Sigma Aldrich), and 1% penicillin/streptomycin mixture. HAEC's were cultured in EBM-2 Basal Medium supplemented with human epidermal growth factor (hEGF), vascular endothelial growth factor (VEGF), R3-insulin-like growth factor-1 (R3-IGF-1), ascorbic acid, hydrocortisone, human fibroblast growth factor-beta (hFGF-β), FBS, and gentamicin/amphotericin-B (GA) (Lonza). Cells were maintained at 37°C in a humidified 5% CO2 incubator. BAE cells and HAEC were used between passages 5–9.
3
Primary HAEC Culture Conditions
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Primary HAECs were cultured in EBM-2 medium–supplemented EGM-2 medium containing hEGF, hydrocortisone, hFGF-b, VEGF, R3-IGF-1, ascorbic acid, Heparin, fetal bovine serum, and Gentamicin/Amphotericin-B [GA] (Lonza). The cells were plated on gelatin-coated plates and kept under a humidified atmosphere of air/CO2 (19:1) at 37°C.
4
Optimizing Myoblast Culture for Fusion
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Normal human myoblasts (MB) were purchased from Lonza Bioscience (Mapleton, IL, USA), and DMD-affected myoblasts were purchased from Creative Bioarray Ltd. (Shirley, NY, USA). Myoblasts (MB) were cultured in Skeletal Muscle Cell Growth Medium-2 (Lonza Clonetics, Mapleton, IL, USA) supplemented with the human Epidermal Growth Factor (hEGF); Fetal Bovine Serum (FBS); Dexamethasone; Gentamicin/Amphotericin B (GA) (Lonza Clonetics, Mapleton, IL, USA). Upon reaching 60–70% confluence, myoblasts were harvested using 0.25% trypsin/EDTA (Sigma-Aldrich, MO, USA). Enzymatic activity was inhibited with 10% serum-supplemented culture media. Human MBs were harvested between passages 3–7, which is optimal for the ex vivo cell fusion procedure.
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