We used previously established Epstein Barr Virus (EBV) transformed LCLs from a NLE (Lazar et al. 2018) (link), and established new LCL for HLE, HMO, and SSY gibbons according to previous protocols (Lazar et al. 2018) (link). Briefly, opportunistic whole blood samples were collected in sodium heparin tubes from each gibbon during routine check-ups at the Gibbon Conservation Center (Santa Clarita, CA). We used Ficoll-Paque PLUS (GE Healthcare) to isolate lymphocytes from the blood. Next, we transformed 3-9 × 106 lymphocytes with EBV from the marmoset cell line B95-8 (ATCC CRL-1612), using a standard protocol. To do so, we incubated the cells with EBV for 2 h at 37 °C, then diluted them with RPMI-1640 (Corning cellgro) supplemented with 10% FBS (Hyclone), 1× MEM Nonessential Amino Acids Solution (Corning cellgro), 1 mM Sodium pyruvate (Corning cellgro) 1% Pen-Strep (Corning cellgro), and 2 mM l -glutamine (Hyclone). Finally, we grew cells undisturbed for 10-12 days and started feeding the cells with the same supplemented RPMI-1640 as soon as signs of transformation were observed under the microscope.
Mem non essential amino acids solution
Manufactured by Corning
MEM non-essential amino acids solution is a cell culture media supplement produced by Corning. It contains a mixture of non-essential amino acids formulated to support cell growth and maintenance in various cell culture applications.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Corning (2 mentions)
Dulbecco modified eagle medium (dmem), by Corning (2 mentions)
Ficoll paque plus, by GE Healthcare (1 mentions)
Rpmi 1640, by Corning (1 mentions)
Sodium pyruvate, by Corning (1 mentions)
Pen strep, by Corning (1 mentions)
3 protocols using mem non essential amino acids solution
1
Establishing Gibbon Lymphoblastoid Cell Lines
2
Culturing Diverse Cell Lines for RBM38 Studies
Check if the same lab product or an alternative is used in the 5 most similar protocols
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MCF7, HCT116, p53−/− HCT116, and H1299 cells were cultured in DMEM (Invitrogen) supplemented with 10% fetal bovine serum (Hyclone) as previously described (Zhang and Chen, 2007). RBM38−/− and p53−/−; RBM38−/− double knockout mouse embryo fibroblasts (MEFs) were generated as described previously 5 (link) and cultured in DMEM supplemented with 10% fetal bovine serum, 55 µM β-mercaptoethanol, and 1× MEM non-essential amino acids solution (Cellgro). MCF7, HCT116, and H1299 cell lines in which RBM38 or HA-tagged RBM38 are inducibly expressed were cultured as previously reported 1 (link), 37 (link). MCF7 cell line in which RBM38 is inducibly knocked down was used as previously reported 1 (link), 37 (link).
3
Culturing Diverse Cell Lines for RBM38 Studies
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