HT22 cells were a generous gift from Dr. David Schubert (The Salk Institute, La Jolla, CA). HT22 cells were fed with Dulbecco’s Modified Eagle Medium (DMEM #11995; Invitrogen/Life Technologies, Grand Island, NY) supplemented with 10% fetal bovine serum (FBS #F4135; Sigma-Aldrich, St. Louis, MO), and cultured at 50–55% relative humidity, in 10% CO2 at 37°C (as recommended by Dr. Schubert). The cells were maintained at ≤50% confluency, as previously described [17 (link)]. It is important to note that sensitivity to glutamate cytotoxicity varies in HT22 neurons depending on their density and passage number. Therefore, experiments were always performed at a constant cell number in early passages of cells. HT22 cells were plated 12 hours before the experiment, and at the beginning of the experiment the entire media was changed in all groups to DMEM + 10% FBS (control); 5mM Glutamate in DMEM + 10% FBS (Glutamate); and 5mM Glutamate + 100 μM tocopherol in DMEM + 10% FBS (Tocopherol treated). L-glutamic acid (#G8415), and (±)-α-Tocopherol (#T3251) were obtained from Sigma-Aldrich (St. Louis, MO).
Fbs f4135
Manufactured by Merck Group
FBS #F4135 is a laboratory equipment product offered by Merck Group. It is a fetal bovine serum, a common cell culture supplement used to support the growth and proliferation of cells in vitro. The product provides a source of essential growth factors, hormones, and other nutrients required for cell culture applications.
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2 protocols using fbs f4135
1
Glutamate-Induced Cytotoxicity in HT22 Cells
2
Generation and Propagation of Stable Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The African green monkey kidney fibroblast cell line (CV-1) was obtained from the American Type Culture Collection (ATCC; CCL-70). The stable cell lines CV-1-pTet-TurboFP635-EF-1a-Egfp and CV-1-EF-1a-TurboFP635 (Figure A1 , Appendix A ) were generated and propagated in Dulbecco’s modified Eagle’s medium (DMEM; 11965092, Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (FBS, F4135, Sigma) and 1% penicillin–streptomycin (P4333, Sigma).
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