LX-2 cells (Merck Millipore; Billerica, MA) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) (Thermo Fisher Scientific; Waltham, MA) supplemented with 2% fetal bovine serum (FBS) and 1% Pen/Strep (Omega Scientific; Tarzana, CA). Approximately 1 x 106 cells were thawed in T-75 flasks (Corning Life Sciences; Corning, NY) containing 12 mL cell culture medium and placed at 37°C in a Hera Cell 5% CO2 incubator (Thermo Fisher Scientific). Culture medium was replaced the first day after thawing, and then every 72 hours until 80% confluent. HepG2 and HEK293 cells (ATCC; Manassas, VA) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) (Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (FBS) and 1% Pen/Strep (Omega Scientific). Culture medium was replaced the first day after thawing, and then every 48 hours until 80% confluent. Primary human hepatocytes (Thermo Fisher Scientific) were thawed in 50 mL Cryopreserved Hepatocyte Recovery Medium (CHRM) and plated in 500 υL William's E Medium supplemented with Hepatocyte Plating Supplement Pack on collagen-coated 24-well plates (Thermo Fisher Scientific). Culture medium was replaced the first day after thawing with William's E Medium supplemented with Hepatocyte Maintenance Supplement Pack.
Hera cell 5 co2 incubator
Manufactured by Thermo Fisher Scientific
The Hera Cell 5% CO2 incubator is a laboratory equipment designed to provide a controlled environment for cell culture applications. It maintains a temperature of 37°C and a 5% CO2 concentration to support the optimal growth and maintenance of cell lines.
Automatically generated - may contain errors
Lab products found in correlation
Lx 2 cells, by Merck Group (2 mentions)
T75 flask, by Corning (2 mentions)
24 well culture dishes, by Avantor (2 mentions)
Primary human hepatocytes, by Thermo Fisher Scientific (1 mentions)
Hepg2 and hek293 cells, by American Type Culture Collection (1 mentions)
Hepatocyte plating supplement pack, by Thermo Fisher Scientific (1 mentions)
Collagen coated 24 well plates, by Thermo Fisher Scientific (1 mentions)
Pen strep, by Omega Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Oil red o dye, by Merck Group (1 mentions)
6 well culture dishes, by Avantor (1 mentions)
Insulin, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Rpmi 1640 medium, by Corning (1 mentions)
Penicillin streptomycin solution, by Corning (1 mentions)
Hepg2, by American Type Culture Collection (1 mentions)
5 protocols using hera cell 5 co2 incubator
1
Cell Culture Protocols for LX-2, HepG2, HEK293, and Primary Hepatocytes
2
BC Cells Culture and Sample Collection
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BC 4T1 cells were purchased from the American Type Culture Collection (ATCC) and cells were cultured in RPMI medium 1640 (GIBCO, Gaithersburg, MD, USA) supplemented with 10% fetal bovine serum (HyClone, Logan, USA) and 1% penicillin/streptomycin (Invitrogen, Waltham, MA). Cells were grown in 24-well culture dishes (VWR International; Radnor, PA) containing 1.0 ml cell culture medium at 37 °C in a Hera Cell 5% CO2 incubator (Thermo Fisher Scientific; Waltham, MA). Culture medium was replaced after 1 day of seeding and then every 48 h thereafter. A total of 60 formalin-fixed paraffin-embedded BC samples were collected from BC patients who underwent curative-intent surgery without prior radiotherapy and chemotherapy at the Department of Pathology of the Third Affiliated Hospital of Southern Medical University. Informed consent was obtained from each patient on the day of admission. The study was approved by the ethics committee of The Third Affiliated Hospital of Southern Medical University.
3
LX-2 Cell Line Characterization
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LX-2 cells (Merck Millipore; Billerica, MA) were cultured in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 2% fetal bovine serum (FBS). Cell line authentication was performed using short tandem repeat (STR) profiling (Cell Line Genetics; Madison, Wi), which confirmed the presence of a single cell line and alleles matching the known DNA fingerprint [44 ]. Cells were grown in T-75 flasks (Corning Life Sciences; Corning, NY) containing 12 mL cell culture medium and placed at 37 °C in a Hera Cell 5% CO2 incubator (Thermo Fisher Scientific; Waltham, MA). Culture medium was replaced the first day after seeding, and then every 72 h. In preparation for treatment with MDI solution, LX-2 cells were seeded at 0.5 × 106 cell/well on 6-well culture dishes (VWR International; Radnor, PA) and serum-starved overnight. Cell culture medium was aspirated and replaced with DMEM, 10% FBS, and MDI solution [0.5 mM isobutylmethylxanthine, 1 μM dexamethasone, and 167 nM insulin (Sigma-Aldrich; St. Louis, MO)] for 72 h to induce a state resembling biological quiescence [45 (link)]. General morphology was observed with a light microscope, lipid accumulation was assessed with Oil Red O dye (Sigma-Aldrich), and levels of alpha smooth muscle actin (ACTA2) were measured by western blot analysis.
4
Establishing Luciferase-Expressing C4-2b-BMP4 Cells
5
Culturing HepG2 Cells in DMEM
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