All tested breast cancer cell lines were obtained from the American Type Culture Collection (Rockville, MD). ER+ cell line MCF-7 and TNBC MDA-MB-231 were maintained in Dulbecco’s modified Eagle’s medium (DMEM) media (Corning Cellgro, NY) supplemented with 10% fetal bovine serum (Corning Cellgro), 1% Penicillin-Streptomycin 100X solution (Corning Cellgro); ER+ cell line T-47D was maintained in RPMI 1640 Medium (Corning Cellgro, NY) supplemented with 10% fetal bovine serum (GIBCO), 1% Penicillin-Streptomycin 100X solution (Corning Cellgro); MCF-10A was maintained in Dulbecco’s Modification of Eagle’s Medium (DMEM)/Ham’s F12 50/50 Mix (Corning Cellgro, NY) supplemented with 10% fetal bovine serum (Corning Cellgro), 1% Penicillin-Streptomycin 100X solution (Corning Cellgro) and 1% ITS (Sigma, I3146) ER −cell line MDA-MB-468 was maintained Leibovitz L-15 Media (HyClone; SH3052501) supplemented with 10% fetal bovine serum (Corning Cellgro), 1% Penicillin-Streptomycin 100X solution (Corning Cellgro); were cultured at 37°C in low CO2 condition (0.5% CO2 : 99.5% air).
Penicillin streptomycin solution 100x
Manufactured by Corning
Sourced in United States
Penicillin-Streptomycin Solution 100X is a sterile, liquid medium supplement used to inhibit the growth of bacteria in cell culture applications. It contains a combination of the antibiotics penicillin and streptomycin at a concentration 100 times greater than standard use. This solution is designed to provide broad-spectrum antimicrobial protection for in vitro cell culture systems.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Merck Group (4 mentions)
Rpmi 1640 medium, by Corning (3 mentions)
Gibco dmem, by Thermo Fisher Scientific (3 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (3 mentions)
D glucose, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Dulbecco modified eagle medium (dmem), by Corning (2 mentions)
Fetal bovine serum (fbs), by Corning (2 mentions)
Fetal bovine serum (fbs), by Gemini Bio (1 mentions)
L glutamine, by Corning (1 mentions)
Mycoalert, by Lonza (1 mentions)
Jeko 1, by American Type Culture Collection (1 mentions)
Z 138, by American Type Culture Collection (1 mentions)
Jvm 2, by American Type Culture Collection (1 mentions)
L ascorbic acid, by Fujifilm (1 mentions)
Mcf 7, by Corning (1 mentions)
Dmem hams f 12 50 50 mix, by Corning (1 mentions)
Dmem media, by Corning (1 mentions)
Phosphate buffered saline (pbs), by Corning (1 mentions)
Glass bottom dish, by Ibidi (1 mentions)
14 protocols using penicillin streptomycin solution 100x
1
Breast Cancer Cell Line Maintenance
2
Maintenance of Mantle Cell Lymphoma Cell Lines
3
Isolation and Culture of Fibroblasts
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All patient-derived fibroblasts were collected with informed written consent from each patient in compliance with the Declaration of Helsinki and the experiments conformed to the principles set out in the Department of Health and Human Services Belmont Report. RDEB patient samples are usually donations from biopsy sites, either of non-SCC or SCC origin, and the non-EB patient samples are from breast reduction skin donations from elective surgeries. Cells were isolated from skin biopsies taken from routine or tumor excision surgeries and cultured at 37 °C at 5% CO2. Primary fibroblasts were grown in Dulbecco’s modified essential medium (DMEM, Corning Cellgro, Mediatech Inc, Manassas, VA) with 10% fetal bovine serum (FBS, PEAK Serum, Cat PS-FB1, Colorado, USA) and 1% Penicillin Streptomycin 100X Solution (Corning, Cat 30-002-CI, Manassas, VA, USA). All media in experimental conditions contained l -ascorbic acid (150 μM, Wako, 012-12061). Non-EB and RDEB fibroblasts (Appendix Table S3 ) were used up to passage 7. Fibroblasts were plated in 6-well plates to achieve 100% confluence within 2 days of seeding, followed by protein extraction or RNA isolation.
4
Visualizing Mitochondrial Dynamics in HeLa Cells
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The penicillin-streptomycin 100x solution, 0.25% Trypsin-EDTA, and Phosphate-Buffered Saline (PBS) were supplied from Corning Cellgro. HyClone Fetal Clone III serum was obtained from GE Healthcare and Modified Eagle’s Medium (MEM) was purchased from Biowest. Carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP) and wt-GFP solution in PBS was purchased from Sigma. Celllight BacMam 2.0 Mito-GFP was provided by Thermofischer.
HeLa cell line was purchased from ECACC (Cat. No. 93021013), grown under standard culture conditions and seeded inside of two silicone inserts wells on a 35 mm glass-bottom dish (Ibidi) in MEM culture media supplemented with 10% serum, 1% of penicillin-streptomycin, incubated under 5% of CO2 at 37 °C. CellLight Mitochondria-GFP BacMam 2.0 was added to the cells in appropriate proportion following the supplier’s protocol and incubated overnight.
HeLa cell line was purchased from ECACC (Cat. No. 93021013), grown under standard culture conditions and seeded inside of two silicone inserts wells on a 35 mm glass-bottom dish (Ibidi) in MEM culture media supplemented with 10% serum, 1% of penicillin-streptomycin, incubated under 5% of CO2 at 37 °C. CellLight Mitochondria-GFP BacMam 2.0 was added to the cells in appropriate proportion following the supplier’s protocol and incubated overnight.
5
Quantification of Drug Metabolism Enzymes
6
Culturing Murine CAD5 Neuronal Cells
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CAD5 cells were a gift from the Prusiner Lab (University of California, San Francisco). CAD5 cells were maintained at 37°C in Opti-MEM I Reduced Serum Medium (Fisher Scientific, Hampton, NH, USA) supplemented with 9% HyClone Bovine Growth Serum (VWR, Radnor, PA, USA) and 100 units Penicillin-Streptomycin Solution, 100x (Corning Inc., Corning, NY, USA). Dividing cells were plated at 10% confluency and were split 1:10 when confluent.
7
Fluorescence Imaging of Cell Lines
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For fluorescence imaging of living cells, human embryonic kidney 293 (HEK-293, ATCC) and human hepatoma cancer cell (HepG2, ATCC) were cultured in Dulbecco′s Modified Eagle′s Medium (DMEM, HyClone) supplemented with 10% Fetal Bovine Serum (FBS, Gibco, Carlsbad, CA, USA) and 1% Penicillin/Streptomycin Solution 100X (Corning, Corning, NY, USA). All cells were cultured in a humidified incubator at 37 °C with 5% CO2. HEK-293 and HepG2 cells, approximately 1 × 104 cells, were seeded on 8-well chambered cover glasses (LabTek, Nunc, Carlsbad, CA, USA) and incubated in complete medium for 24 h. After that, the cells were treated with 5 μM stock solutions of 3a –f in DMSO/cell culture medium containing 10% FBS (0.25% DMSO), for 0, 1, 3, and 6 h. Then, the cells were washed twice with 0.01 M PBS before being visualized under a Laser-Scanning Confocal Microscope (Nikon A1Rsi) with a 60× oil immersion objective lens and living cell workstation with the excitation channel at 488 nm.
8
Breast Cancer Cell Line Characterization
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Human breast cancer cell lines MCF7 and ZR75-1 were obtained from the American Type Culture Collection (ATCC, USA). Cells were cultured in DMEM (Dulbecco's modified Eagle's medium, Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (Gibco, Carlsbad, CA, USA) and 1% penicillin-streptomycin solution 100X (Corning, CA, USA) in a humidified atmosphere with 5% CO2 at 37 °C. Specific antibodies against PGM5 (PA5-48880) was obtained from Invitrogen. Anti-p21 (ab227443), anti-p53 (ab131442), anti-BAX (ab216494), anti-FAK (ab40794), and anti-LDHB (ab112996) were purchased from Abcam. Antibodies against cyclin B (#4138), cyclin D1 (#2922), Bcl-2 (#2876), phos-FAK (Y397) (#8556), MMP-2 (#4022), MMP-9 (#3852), and LDHA (#3582) were obtained from Cell Signaling. Anti-E-cadherin (sc-8426), anti-vimentin (sc-6260), anti-N-cadherin (sc-8424), and anti-β-actin (sc-47778HRP) antibodies were purchased from Santa Cruz Biotechnology.
9
Culture of Immortalized Mouse Spermatogonia
10
MA-10 Leydig Cell Culture Protocol
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