NIH/3T3 mouse fibroblast cells (ATCC) were grown in DMEM supplemented with 10% calf serum, sodium pyruvate and penicillin/streptomycin (Invitrogen) at 37 °C, 5% CO2. MDA-MB-231 breast cancer cells (ATCC) were grown in Leibovitz’s L15 media supplemented with 10% fetal bovine serum and penicillin/streptomycin at 37 °C in atmospheric air. Cell lines have been tested yearly for mycoplasma and are free of contamination.
Mda mb 231 breast cancer cells
Manufactured by American Type Culture Collection
Sourced in United States
MDA-MB-231 breast cancer cells are a well-established cell line derived from a human breast adenocarcinoma. They are commonly used in cancer research.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (12 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (9 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (6 mentions)
Penicillin, by Thermo Fisher Scientific (4 mentions)
Streptomycin, by Thermo Fisher Scientific (3 mentions)
Hs 5 and hs 27a bone marrow stromal cells, by American Type Culture Collection (2 mentions)
Hyclone, by GE Healthcare (2 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (2 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Corning (2 mentions)
Hek293t cells, by Thermo Fisher Scientific (2 mentions)
Dmem f12, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Penicillin streptomycin, by Merck Group (2 mentions)
Hct116 colon cancer cells, by American Type Culture Collection (2 mentions)
Skov 3 ovarian cancer cells, by American Type Culture Collection (2 mentions)
Hepes, by Merck Group (2 mentions)
Insulin, by Merck Group (2 mentions)
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Hydrocortisone, by Merck Group (2 mentions)
59 protocols using mda mb 231 breast cancer cells
1
Cell Culture Protocol: NIH/3T3 and MDA-MB-231
2
Co-culture of Breast Cancer Cells and ASCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For co-culture experiments, hydrogels containing MDA-MB-231 breast cancer cells (ATCC, Manassas, VA, USA) were printed on top of the ASC-laden constructs after 21 days of adipogenic culture. MDA-MB-231 breast cancer cells were expanded in growth medium composed of DMEM (1 g/L glucose, Life Technologies) supplemented with 10% FBS, 1% L-glutamine, 1% MEM non-essential amino acids solution, 1% sodium pyruvate, 0.6% HEPES, and 1% penicillin/streptomycin (all Life Technologies). After harvesting, MDA-MB-231 cells at passage 6 were suspended in hyaluronic acid-based hydrogel precursor solution and printed at 1 bar as described above. Before transferring the suspension into the glass rings for UV irradiation (see above), differentiated ASC spheroid-laden hydrogels were put at the bottom inside the glass ring. The hydrogel–cancer cell suspension was then dispensed on top and UV-crosslinked at 365 nm for 10 min. The resulting co-culture hydrogels (bottom: adipogenically differentiated ASC spheroids, top: MDA-MB-231 cells) were cultured for 9 days in maintenance medium consisting of DMEM/F12, 10% fetal bovine serum, 1.7 µM insulin, and 1% penicillin/streptomycin. Samples were harvested at d0 and d9. Double gels with either empty top or bottom compartment (only ASC spheroids at the bottom or only MDA-MB-231 at the top) served as monoculture controls.
3
Culturing Diverse Cancer Cell Lines
4
Cell Culture Conditions for HT1080 and MDA-MB-231
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HT1080 fibrosarcoma cells (ATCC) were cultured in Eagle’s mimimum essential medium and MDA-MB-231 breast cancer cells (ATCC) were cultured in Dulbecco’s modified essential medium. Culture media were supplemented with 10% heat-inactivated FBS and 40 μg ml−1 gentamicin. Cells were cultured in a humidified atmosphere at 37 °C with 5% CO2 and 21% O2. For incubation under hypoxic conditions, cells were placed in an In Vivo2 400 hypoxia workstation (Ruskinn) under a humidified atmosphere of 1% O2 and 5% CO2. All cell lines were routinely tested for mycoplasma using the MycoSEQ Mycoplasma Detection Kit (Thermo Fisher Scientific).
5
Cell Culture Conditions for Microscopy
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We cultured HEK 293T cells (Thermo Fisher Scientific, Carlsbad, CA), MDA-MB-231 breast cancer cells (ATCC), and HS-5 and HS-27A bone marrow stromal cells (ATCC) in Dulbecco's modified eagle's medium (Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (FBS) (HyClone; GE HealthCare, Logan, UT). For microscopy, we replaced the culture medium with phenol red-free DMEM (Thermo Fisher Scientific) supplemented with 1% FBS, 0.1 nM estrogen, 1% penicillin/streptomycin, 1% pyruvate, and 2.5% glutamine unless stated otherwise. Cells were cultured at 37°C in an incubator with 5% CO2.
6
Cell Culture and Maintenance Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DU145 prostate cancer (PCa) cells were purchased from ATCC (Manassas, VA). PPC1 PCa cells were originally obtained from Dr. Arthur Brothman [51 (link)]. Cells were maintained in 10% fetal bovine serum (FBS) (Gemini, West Sacramento, CA) RPMI (Cellgro, Manassas, VA). MDA MB 231 breast cancer cells were obtained from ATCC and cultured in DMEM (Cellgro) supplemented with 10% FBS. All cells were maintained in a 37°C incubator with 5% CO2. Cells were subcultured upon attaining 75% confluence.
7
Culturing Breast Cancer Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MDA-MB-231 breast cancer cells (ATCC,
Manassas, VA, USA) and MCF-7 mammary tumor cells (ATCC, Manassas,
VA, USA) lentivirally transduced to constitutively express firefly
luciferase were cultured on standard tissue culture-treated polystyrene
in an incubator maintained at 37 °C and 5% CO2. Cells
were maintained in growth media consisting of Dulbecco’s modified
Eagle’s medium (DMEM; Gibco-Life Technology) supplemented with
10% fetal bovine serum (FBS; Gibco) and 50 μg/mL gentamicin
(Mediatech, Englewood, CO, USA).
Manassas, VA, USA) and MCF-7 mammary tumor cells (ATCC, Manassas,
VA, USA) lentivirally transduced to constitutively express firefly
luciferase were cultured on standard tissue culture-treated polystyrene
in an incubator maintained at 37 °C and 5% CO2. Cells
were maintained in growth media consisting of Dulbecco’s modified
Eagle’s medium (DMEM; Gibco-Life Technology) supplemented with
10% fetal bovine serum (FBS; Gibco) and 50 μg/mL gentamicin
(Mediatech, Englewood, CO, USA).
8
MDA-MB 231 Breast Cancer Cell Viability Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MDA-MB 231 breast
cancer cells were purchased from ATCC (Manassas, VA). Cell culture
media and reagents were purchased from Invitrogen. 3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium
(MTS)-cell viability assay reagents were purchased from Promega. Cells
were cultured in high glucose DMEM containing 10% fetal bovine serum.
Cells were plated in 96-well microtiter plates at 5000 cells per well
and incubated for 2 h to allow for adherence. Where appropriate, pH
7.4 media was exchanged for pH 6.0 media adjusted with 1 M HCl to
determine PA cytotoxicity at acidic pH. After 2 h, 10 μL of
PA was added for the final appropriate concentrations. Cell viability
was measured after 48 h using an MTS cell viability assay and was
used according to the supplier’s instructions. Briefly, the
cell media was replaced after 48 h with a stock of 20% MTS solution
in pH 7.4 media. The plate was incubated for 1–3 h, and absorbance
was read using a Molecular Devices microplate reader (490 nm). Cell
viability was calculated as an absorbance percent relative to the
untreated cell control at the same pH. Experiments were performed
in triplicate.
cancer cells were purchased from ATCC (Manassas, VA). Cell culture
media and reagents were purchased from Invitrogen. 3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium
(MTS)-cell viability assay reagents were purchased from Promega. Cells
were cultured in high glucose DMEM containing 10% fetal bovine serum.
Cells were plated in 96-well microtiter plates at 5000 cells per well
and incubated for 2 h to allow for adherence. Where appropriate, pH
7.4 media was exchanged for pH 6.0 media adjusted with 1 M HCl to
determine PA cytotoxicity at acidic pH. After 2 h, 10 μL of
PA was added for the final appropriate concentrations. Cell viability
was measured after 48 h using an MTS cell viability assay and was
used according to the supplier’s instructions. Briefly, the
cell media was replaced after 48 h with a stock of 20% MTS solution
in pH 7.4 media. The plate was incubated for 1–3 h, and absorbance
was read using a Molecular Devices microplate reader (490 nm). Cell
viability was calculated as an absorbance percent relative to the
untreated cell control at the same pH. Experiments were performed
in triplicate.
9
Cultivation of Human Dermal and Mammary Fibroblasts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Normal human dermal fibroblasts (NHDFs) and Fibroblast Growth Medium containing 2% fetal calf serum (FGM), insulin (5μg/mL) and FGF-2 (1ng/mL) were obtained from PromoCell (http://www.promocell.com/ ) and cells were cultured as specified by the supplier. Human Primary Mammary Fibroblasts (HPMFs), HPMF growth media (HPMF-GM), the Fibroblast Medium Supplement Kit (FBS, hydrocortisone, L-glutamine, FGF and an antibiotic-anti-mycotic solution) and gelatin coating solution were obtained from Cell Biologics (http://www.cellbiologics.net ). HPMF cells were cultured in HPMF-GM with the added supplements for 6–7 passages as specified by the supplier. MDA-MB-231 breast cancer cells were obtained from ATCC and cultured for 12–15 passages in MCF media. MCF medium is DMEM/F12 (Life Technologies, Carlsbad, CA) supplemented with 5% horse serum, human recombinant EGF, insulin, hydrocortisone and cholera toxin, all obtained from Sigma Aldrich (St. Louis, MO), and penicillin-streptomycin from Thermo -Fisher (Grand Island, NY) [21 (link)]. GFP tagged human dermal fibroblasts (NHDFs-GFP) were obtained from Angio-proteomie (www.angioproteomie.com ) and cultured according to the supplier’s directions.
10
Intracellular Localization of Quantum Dots
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For intracellular localization of QDs, QD-Ce6 complexes and Ce6, MDA-MB-231 breast cancer cells (ATCC, USA) were seeded on a Nunc LabTek II chamber slide (Thermo Fisher Scientific, Waltham, MA, USA) at a density of 25,000 cells per well using standard cell culture medium (Dulbecco’s modified Eagle’s medium (DMEM), 10% fetal bovine serum (FBS), 1% penicillin/streptomycin antibiotic mix (all from Gibco, Waltham, MA, USA). Upon cell attachment, cells were treated with serum-free cell medium containing 10 nM of either different P-QDs or L-QDs or their QD-Ce6 complexes for 24 hours. In addition, 1 μM Ce6 solution was used as a control. Samples for confocal microscopy were prepared as previously described [22 (link)]. Briefly, cells were fixed with 4% paraformaldehyde solution (Sigma Aldrich, USA) and permeabilized with 0.2% Triton-X 100 (Sigma Aldrich, St. Louis, MO, USA). Actin filaments were stained with 15 U/mL Alexa Fluor 488 Phalloidin (Thermo Fisher Scientific, Waltham, MA, USA) and 25 μg/mL Hoechst 33342 (Sigma Aldrich, St. Louis, MO, USA) was used to label nuclei.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!