Human liver cancer HepG2 cell line (ATCC HB8065) and rat cardiomyoblast cell line H9c2 (ATCC-CRL1446) were purchased in ATCC (American Type Culture Collection, Manassas, VA, USA). The cells were cultured in standard conditions (37 °C, 5% CO2) in EMEM medium (ATCC, Manassas, VA, USA), supplemented with 10% FBS (Gibco, Waltham, MA, USA) and antibiotics (1% streptomycin/penicillin mixture, Sigma–Aldrich, Darmstadt, Germany) or DMEM high glucose (ATCC, Manassas, VA, USA) supplemented with 10% FBS (Gibco, Waltham, MA, USA) and antibiotics (1% streptomycin/penicillin mixture, Sigma–Aldrich, Darmstadt, Germany), respectively.
Emem medium
Manufactured by American Type Culture Collection
Sourced in United States
EMEM medium is a commonly used cell culture medium that provides nutrients and growth factors necessary for the maintenance and proliferation of a variety of cell types. It is a balanced salt solution supplemented with amino acids, vitamins, and other essential components to support cell growth and viability.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (22 mentions)
Hepg2, by American Type Culture Collection (8 mentions)
Dmem medium, by Thermo Fisher Scientific (6 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (5 mentions)
Rpmi 1640 medium, by American Type Culture Collection (5 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (4 mentions)
Fetal bovine serum (fbs), by American Type Culture Collection (4 mentions)
Plc prf 5, by American Type Culture Collection (4 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (3 mentions)
Hep3b, by American Type Culture Collection (3 mentions)
Pen strep, by Thermo Fisher Scientific (3 mentions)
Rpmi 1640 medium, by Merck Group (3 mentions)
Fetal bovine serum (fbs), by Gemini Bio (2 mentions)
Bovine insulin, by Merck Group (2 mentions)
β estradiol, by Merck Group (2 mentions)
Alexander plc prf 5, by American Type Culture Collection (2 mentions)
Dulbecco modified eagle medium (dmem), by American Type Culture Collection (2 mentions)
Hpaf 2, by American Type Culture Collection (2 mentions)
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Mycoalert mycoplasma detection kit, by Lonza (2 mentions)
65 protocols using emem medium
1
Cell Culture of Liver Cancer and Cardiomyoblast Lines
2
Mammalian Cell Culture and Transfection
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The human hepatocellular carcinoma cell line HepG2 (ATCC HB-8065) was maintained in EMEM medium (ATCC) supplemented with 10% FBS (Invitrogen, Carlsbad, CA), 2 mM L-Glutamine and 1% penicillin-streptomycin. HEK293T and Hela cells were grown in DMEM medium (Invitrogen) containing 10% FBS, 2 mM L-Glutamine and 1% penicillin-streptomycin. All cell lines were purchased from the American Type Culture Collection (ATCC) and were not authenticated by ourselves. All cell lines were routinely tested for mycoplasma contamination. These cell lines were not listed in the database of commonly misidentified cell lines maintained by ICLAC. For heat shock treatment, Hela cells were incubated at 42 °C for 1 hour. Plasmids and siRNAs were transfected into cells with Lipofectamine 2000 (Invitrogen) according to the instructions of the manufacturer.
3
HepG2 Cell Culture Protocol
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Human hepatic cell line HepG2 was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA), and cells were cultured in EMEM medium (ATCC) supplemented with 10% foetal calf serum (Gibco, Thermo Fisher Scientific, Waltham, MA, USA). The cells were incubated in 5% CO2 atmosphere at 37 °C. Asynchronous cell cultures in the exponential phase of growth were used in all experiments.
4
Cell Culture and BMP Treatment Protocol
5
Culturing Human Neuroblastoma Cell Lines
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Human neuroblastoma cell lines SK-N-BE (2) and SK-N-SH were purchased from the American Type Culture Collection (ATCC, USA). All cells were cultured in EMEM medium (ATCC, USA) containing 10% of fetal bovine serum (FBS; Gibco, USA), in an incubator at 37°C with 95% humidity and 5% CO2.
6
Retinoic Acid Differentiation of SK-N-SH Cells
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SK-N-SH cells were purchased from the American Type Culture Collection (ATCC) and cultured using EMEM medium (ATCC) complemented with 10% FBS (Thermo Fisher Scientific). Cells were differentiated for 48 h using 6 μM of all-trans retinoic acid (Sigma-Aldrich).
After harvesting, total RNA were extracted using Tripure isolation reagent (Sigma-Aldrich), treated with DNase I (DNAfree, Ambion) for 30 min at 37 °C and reverse-transcribed (RT) using M-MLV reverse transcriptase and random primers (Invitrogen). Before PCR, all RT reaction mixtures were diluted at 2.5 ng μL of initial RNA. PCR reactions were performed using GoTaq polymerase (Promega).
MCF7 cells were cultured as described in36 . RT-PCRs were performed using the same protocol as for SK-N-SH cells.
After harvesting, total RNA were extracted using Tripure isolation reagent (Sigma-Aldrich), treated with DNase I (DNAfree, Ambion) for 30 min at 37 °C and reverse-transcribed (RT) using M-MLV reverse transcriptase and random primers (Invitrogen). Before PCR, all RT reaction mixtures were diluted at 2.5 ng μL of initial RNA. PCR reactions were performed using GoTaq polymerase (Promega).
MCF7 cells were cultured as described in36 . RT-PCRs were performed using the same protocol as for SK-N-SH cells.
7
Cytotoxicity Assay of Compound in Human Fibroblasts
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ATCC CCL-110 cell line (normal, human fibroblasts derived from skin) was used in the experiment. Cells were cultured in EMEM medium (ATCC) and seeded in 96-wells plate for 24 h. Incubation with tested compound at the chosen concentrations: 0.109, 0.218, 0.437, 0.655, 0.873 µM and 1.091 µM (compound was dissolved in methanol) lasted 48 h. Final methanol concentration in each well was 1 µL per 100 µL of medium. EMEM medium with the addition of 1 µL methanol was used as a control and a positive control was pure methanol. After 48 h of incubation, the cytotoxicity test was carried out according to Vybrant MTT Cell Proliferation Assay Kit: 10 µL of the 12 mM MTT stock solution was added to the cells and further incubated at 37 °C for 4 h. After incubation, not all medium but 25 µL was removed from the wells and 50 µL of DMSO was added. Plates were incubated for 10 min at 37 °C and absorbance at 540 nm was read [27 (link),28 (link),29 (link)].
8
Breast Cancer Cell Line Culture
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SUM159 and SUM149 BC cells were cultured in Ham’s F-12 (ThermoFisher Scientific) supplemented with 5% FBS (ThermoFisher Scientific), 5 µg/mL insulin (Sigma-Aldrich, St. Louis, MO), 1 µg/mL hydrocortisone (Sigma-Aldrich), and 1x antibiotic-antimycotic (ThermoFisher Scientific, 100x). MCF7 cells were grown in EMEM medium (ATCC) supplemented with 10% FBS, 1x antibiotic-antimycotic, and 10 µg/mL insulin (Sigma-Aldrich). BT20, MDA-MB-231, MDA-MB-468, MDA-MB-157 and SKBR3 were cultured in DMEM-high glucose (Gibco) supplemented with 10% FBS and 1x antibiotic-antimycotic. Vari068, HCC38, HCC70, HCC1937, HCC1954, T47D, ZR-75-1 and BT474 were maintained in RPMI1640 medium (ThermoFisher Scientific) supplemented with 10% FBS and 1x antibiotic-antimycotic. MCF10A is cultured in DMEM/F12 media (50:50, ThermoFisher Scientific) supplemented with 5% horse serum, 1x HEPES, 20 ng/ml EGF, 0.5 mg/ml hydrocortisone, 100 ng/ml cholera toxin, 10 µg/ml insulin and 1x antibiotic-antimycotic. All the cell lines are cultured at 37 °C under 5% CO2 in a humidified chamber and are mycoplasma-free.
9
Cell Culture and RNA Extraction Protocol
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HEK293T, HeLa, and MCF7 cells were cultured under standard conditions. Briefly, HEK293T and HeLa cells were grown in Hyclone DMEM medium (GE Healthcare Life Sciences, SH30022.01) with 10% FBS and 1% Pen–Strep (Penicillin–Streptomycin) to 80% confluency. MCF7 cells were grown in EMEM medium (ATCC, 30-2003) with 10% FBS, 1% Pen–Strep, 0.01 mg/mL bovine insulin (Sigma-Aldrich, I0516), and 10 nM β-estradiol (Sigma-Aldrich, E2758) to 80% confluency. Total RNA was extracted using TRIzol reagent (ThermoFisher, 15596026) following the manufacturer's protocol. Poly(A)+ RNA was enriched using the PolyATtract mRNA Isolation System (Promega, Z5310) following the manufacturer's instructions.
10
Culturing Human Amnion Cells
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WISH human amnion cells were purchased from the American Type Culture Collection (CCL25; ATCC, Manassas, VA, USA). The cells were cultured in EMEM medium (30-2003; ATCC) supplemented with 10% fetal bovine serum (Gibco, Carlsbad, CA, USA) in a 5% CO2 atmosphere at 37℃. Three days later, adherent cells were removed and the culture was continued, with replacement of the medium twice a week.
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