MCF7 (ATTC©HTB-22) BC cell line was obtained from the American Type Culture Collection (Manassas, VA, USA). BC cells were grown in DMEM/HAM F12 culture medium (Sigma, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (FBS) (Sigma; St. Louis, MO, USA), 100 IU/mL of penicillin (Sigma; St. Louis, MO, USA), and 100 µg/mL of streptomycin (Sigma; St. Louis, MO, USA). Mycoplasma-free cultures were maintained at 37 °C in a humidified atmosphere with 5% CO2.
Dmem ham f12 culture medium
Manufactured by Merck Group
Sourced in United States
DMEM/HAM F12 culture medium is a commonly used cell culture medium that provides a balanced formulation of nutrients, vitamins, and salts to support the growth and maintenance of various cell lines. It is a combination of Dulbecco's Modified Eagle's Medium (DMEM) and Ham's F12 medium, designed to create a versatile and widely applicable cell culture environment.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Merck Group (3 mentions)
Streptomycin, by Merck Group (3 mentions)
Mcf 7, by American Type Culture Collection (2 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Harvard Bioscience (1 mentions)
Khco3, by Merck Group (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Nh4cl, by Merck Group (1 mentions)
Mda mb 231, by American Type Culture Collection (1 mentions)
Mcf 10a, by American Type Culture Collection (1 mentions)
Bt549, by American Type Culture Collection (1 mentions)
Mda mb 468, by American Type Culture Collection (1 mentions)
Hcc1937, by American Type Culture Collection (1 mentions)
4 protocols using dmem ham f12 culture medium
1
MCF7 Breast Cancer Cell Culture
2
Isolation and Cultivation of Adipocyte Precursor Cells
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Subcutaneous AT samples were obtained from children undergoing bariatric or elective orthopedic surgery (Supplementary Table 1 ). SVF cells and adipocytes were isolated and preserved for later RNA isolation as previously described19 (link),46 . The remaining SVF cells were filtered through a nylon mesh with 30-µm pore size. Erythrocytes were removed with erythrocyte lysis buffer (0.154 M NH4Cl (Sigma-Aldrich), 0.01 M KHCO3 (Merck) and 0.1 mM EDTA (Sigma-Aldrich)). SVF cells were frozen in liquid nitrogen in Dulbecco’s modified Eagle medium/Ham F-12 culture medium (DMEM/F-12) (Life Technologies) containing 10% fetal bovine serum (FBS) (Biochrom) and 10% dimethyl sulfoxide (DMSO) (Sigma-Aldrich). Cells were thawed and 24 h after seeding cells were washed three times with phosphate-buffered saline to select for adipocyte precursor cells via plastic adherence. Isolated SVF cells were cultivated in DMEM/Ham F-12 culture medium containing 10% FBS and 100 U penicillin and 0.1 mg ml−1 streptomycin (Sigma-Aldrich) at 37 °C and 5% CO2 and passaged every 3–4 d.
3
Culturing MDA-MB-231 Breast Cancer Cells
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MDA-MB-231 (ATTC©HTB-26TB) breast cancer cell line was obtained from the American Type Culture Collection (Manassas, VA, USA). Breast cancer cells were grown in DMEM/HAM F12 culture medium (Sigma, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (FBS) (Sigma), 100 IU/mL of penicillin (Sigma), and 100 µg/mL of streptomycin (Sigma). Mycoplasma free cultures were maintained at 37 °C in a humidified atmosphere with 5% CO2.
4
Breast Cancer Cell Line Characterization
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The T47D (ATCC©HTB-133™), MCF7 (ATCC©HTB-22™), MDA-MB-231 (ATTC©HTB-26TM), MDA-MB-468 (ATTC©HTB-132TM), BT-549 (ATTC©HTB-122TM) and HCC1937 (ATTC©CRL-2336TM) BC cell lines as well as MCF-10A (ATTC©CRL-10317TM) normal breast cells were obtained from the American Type Culture Collection (ATTC) (Manassas; VA, USA). Normal human primary fibroblast culture (HSF) was obtained by the outgrowth technique from skin explants of a young person using a method routinely used in our lab (local ethics committee permission No. KE0254/298/2015). BC cells were maintained in the DMEM/HAM F12 culture medium (Sigma-Aldrich). The medium comprised 10% FBS and antibiotics: penicillin—100 IU/mL, streptomycin—100 µg/mL. The cancer cell lines for the research were chosen to evaluate the differences in responses to test substances. The selected cell lines used in our experiments are models of a specified molecular subtype of BC which makes the obtained results much more reproducible and comparable with other studies. The detailed characteristics of BC cell lines used in the study are summed up in Table 1 .
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