Primary human adipose-derived stem cells (hADSCs) (Lonza) were cultured in hADSC basal medium (Lonza) supplemented with 10% fetal bovine serum (FBS), 5 ml L-glutamine (Lonza) and 0.5 ml gentamicin-amphotericin (Lonza) at 37 °C in humidified atmosphere with 5% CO2. Medium was changed every other day. Cells at ~80% confluence were detached for passaging and/or further experimental use. Only cells at or before passage 5 were used.
Gentamicin amphotericin
Manufactured by Lonza
Sourced in United States, Switzerland
Gentamicin/amphotericin is a lab equipment product used for the cultivation and analysis of microorganisms. It is a mixture of the antibiotics gentamicin and amphotericin, which are used to inhibit the growth of bacteria and fungi, respectively. This product is commonly used in cell culture and microbiology research.
Automatically generated - may contain errors
Lab products found in correlation
Hydrocortisone, by Lonza (3 mentions)
Rock inhibitor y 27632, by STEMCELL (2 mentions)
Glutamax, by Thermo Fisher Scientific (2 mentions)
Epidermal growth factor (egf), by Corning (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Heparin, by Lonza (2 mentions)
Penicillin streptomycin solution, by Thermo Fisher Scientific (2 mentions)
Recombinant human fb growth factor, by Lonza (2 mentions)
Insulin like growth factor 1, by Lonza (2 mentions)
Hadscs, by Lonza (1 mentions)
Hadsc basal medium, by Lonza (1 mentions)
L glutamine, by Lonza (1 mentions)
Bxpc 3, by American Type Culture Collection (1 mentions)
Beas 2b, by Thermo Fisher Scientific (1 mentions)
Lhc 8 media, by Thermo Fisher Scientific (1 mentions)
Ultra low attachment 96 well plate, by Corning (1 mentions)
Matrigel, by BD (1 mentions)
Celltiter glo 3d viability assay, by Promega (1 mentions)
Dimethyl sulfoxide (dmso), by Selleck Chemicals (1 mentions)
Nci h660 cells, by American Type Culture Collection (1 mentions)
7 protocols using gentamicin amphotericin
1
Culturing Primary Human Adipose-Derived Stem Cells
2
Cell Line Culture Protocols
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All cell lines were purchased from the American Type Culture Collection (ATCC) (except HMLE and BxPC-3) and all tested negative for mycoplasma. A549 (CCL-185, male), NMuMG (CRL-1636, female) and MCF-7 (HTB-22, female) cells were supplemented with Dulbecco’s Modified Eagle’s Media (DMEM) with 10% fetal bovine serum (Hyclone). BEAS-2B (CRL-9609, male) were supplemented with LHC-8 media (Thermo-fisher scientific) with and without FBS (Hyclone, Canada, characterized). Panc 10.05 (CRL-2547, male) and BxPC-3 cells (CRL-1687, female), a generous gift from Dr. David Hoskin, Dalhousie University, were supplemented with Roswell Park Memorial Institute (RPMI) with 10% fetal bovine serum. HMLE (human mammary epithelial cell line, female) cells were a generous gift from Dr. Robert Weinberg (Whitehead Institute for Biomedical Research, Cambridge, Massachusetts) and were cultured in a 1:1 ratio of DMEM F12 1:1 and mammary epithelial cell growth medium (MEGM, Lonza) supplemented with 13 µg/mL bovine pituitary extract, 20 µg/mL human epidermal growth factor,10 µg/mL insulin, 1 µg/mL gentamicin/amphotericin and 2 µg/mL hydrocortisone (Lonza Clonetics) and 10% FBS. All cells were cultured in the presence of 1% pencillin/streptomycin (Hyclone) and were maintained at 37 °C with 5% CO2.
3
EPHB4 Inhibitor Treatment of Organoid Cultures
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NCI-H660 cells (ATCC) were seeded at 5000 cells/well density in ultra-low attachment 96-well plates (Corning) and cultured in Hepatocyte growth media (Corning) supplemented with 10 ng/ml epidermal growth factor (Corning), 5% heat inactivated charcoal stripped FBS, 1X Glutamax (Gibco), 5% Matrigel (BD Biosciences), 10 µM ROCK inhibitor (Y-27632, STEMCELL Technologies), and 1X Gentamicin/Amphotericin (Lonza), as described previously55 (link). At day 8, organoids were treated with NVP-BHG 712 (EPHB4i) from Selleck Chemicals or DMSO (Veh.), and viability was assessed at day 15 as per manufacturer’s protocol (Cell Titer Glo-3D viability assay, Promega).
4
Culturing Human Pulmonary Endothelial and Smooth Muscle Cells
5
Cell Culture Protocols for Skin Tissue Research
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We used HaCaT cells, HS27 fibroblasts (FBS), and HUVECs as substitutes for the epithelial cells, FBS and blood vessel endothelial cells present in skin tissue54 (link). The human epidermal keratinocytes (HaCaT, ATCC®, USA) and human skin Fb HS27 cells (CRL-1634, ATCC®, USA) were cultured in high-glucose DMEM (WELGENE, South Korea) supplemented with 10% fetal bovine serum (FBS, Gibco, Grand Island, NY) and 1% penicillin/streptomycin solution (Gibco, Grand Island, NY). The HUVECs (PCS-100-010™, ATCC®, USA) were cultured in EGM-2 supplemented with ascorbic acid, vascular endothelial growth factor, 20% FBS, recombinant human Fb growth factor, hydrocortisone, insulin-like growth factor-1, a recombinant analog of human insulin-like growth factor-1, gentamicin–amphotericin, and heparin (Lonza, Walkersville, MD, USA). All experiments used cells from passages 3–10.
6
Culturing Skin Cell Lines for Research
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Human epidermal keratinocytes HaCaT (Addexbio, San Diego, CA, USA), a spontaneously transformed immortal human keratinocyte cell line, and human skin fibroblasts Hs27 (CRL1735, ATCC®, Manassas, VA, USA) were cultured in high-glucose DMEM (WELGENE, Daegu, Korea) supplemented with 10% fetal bovine serum (Gibco, Grand Island, NY, USA) and 1% penicillin/streptomycin solution (Gibco). The HUVECs (PCS-100-01, ATCC®) were cultured in EGM-2 supplemented with ascorbic acid, vascular endothelial growth factor, 20% FBS, recombinant human Fb growth factor, hydrocortisone, insulin-like growth factor-1, a recombinant analog of human insulin-like growth factor-1, gentamicin–amphotericin, and heparin (Lonza, Basel, Switzerland). HaCaT, Hs27, and HUVECs were used as substitutes for epithelial cells, fibroblasts, and blood vessel endothelial cells present in the skin tissue. All experiments used cells from passages 3–10. All cells were incubated in 5% CO2 at 37 °C.
7
Hepatocyte Culture Optimization Protocol
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Cells were resuspended in Hepatocyte Defined Medium (Corning) supplemented with 10 ng/ml epidermal growth factor (Corning), 5% C.S. FBS, 1× Glutamax (Gibco), 5% matrigel (Corning), 10 µM ROCK inhibitor (Y-27632, STEMCELL Technologies), 100 nM DHT (Sigma), and 1× Gentamicin/Amphotericin (Lonza). Cells were plated in Ultra-Low Attachment Surface plates (Corning).
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