Human cell lines HA1800, U87, T98G, U118, A172, U251, and HMC3 were purchased from ScienCell Research Laboratory, Cell Bank of the Chinese Academy of Sciences and ATCC, and cultured in Dulbecco’s modified Eagle’s medium (DMEM) medium (Invitrogen, Thermo Fisher Scientific, Inc., the United States). All the culture media was supplement by 10% fetal bovine serum (FBS) (Gibco), Penstrep (Gibco), GlutaMAX (Gibco), and MEM non-Essential Amino Acids (MEM-NEAA) (Gibco) following the instruction of the manufacturer. These cells were all cultured in an incubator with 5% CO2 at 37°C.
Mem nonessential amino acids mem neaa
Manufactured by Thermo Fisher Scientific
Sourced in United States
MEM nonessential amino acids (MEM NEAA) is a cell culture supplement that provides a mixture of nonessential amino acids. It is designed to be added to cell culture media to support cell growth and development.
Automatically generated - may contain errors
Lab products found in correlation
Glutamax, by Thermo Fisher Scientific (3 mentions)
Esgro lif, by Merck Group (2 mentions)
Knockout dmem with 4.5 g l d glucose, by Thermo Fisher Scientific (2 mentions)
2 mercaptoethanol bme, by Thermo Fisher Scientific (2 mentions)
Knockout serum replacement ksr, by Thermo Fisher Scientific (2 mentions)
Pen strep, by Thermo Fisher Scientific (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Ha1800, by ScienCell (1 mentions)
Irradiated mouse embryonic fibroblasts, by R&D Systems (1 mentions)
C57bl 6j, by Jackson ImmunoResearch (1 mentions)
3 protocols using mem nonessential amino acids mem neaa
1
Culturing Human Cell Lines
2
Establishing Pluripotent Stem Cell Lines
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In-house stem cell expression profiling experiments used material as follows. A 129S6/SvEv embryonic stem cell line was obtained from Millipore Sigma, (Catalog no. SCR012, Burlington, MA, USA). The hybrid embryonic stem cell line was 129Cas (see above). To establish C57BL/6 J and CAST/EiJ induced pluripotent stem cell lines, we used mouse embryonic fibroblasts E13.5 obtained from Jackson Laboratories (Bar Harbor, ME, USA) as input into a stem cell derivation protocol as previously described (Terzic et al. 2016) (link). Briefly, we used octamer-binding transcription factor-4 (Oct4), sex-determining region y-box 2 (Sox2), and Kruppel-like factor-4 (Klf4) as reprogramming factors, introduced using pMXs retroviral vectors.
Stem cells were cultured on irradiated mouse embryonic fibroblasts (R and D Systems, Minneapolis, MN, USA) in miPSC medium: knockout DMEM with 4.5 g/ L d-glucose (Gibco, Grand Island, NY, USA), 10% knockout serum replacement (KSR) (Gibco), 10% fetal bovine serum (FBS) (HyClone, Logan, UT, USA), 1× MEM nonessential amino acids (MEM NEAA) (Gibco), 1× GlutaMAX (Gibco), 0.1 mM 2-mercaptoethanol (BME) (Life Technologies, Grand Island, NY, USA), and 0.02% ESGRO-LIF (Millipore, Billerica, MA, USA). Cells were incubated at 37 °C in 5% CO 2 .
Stem cells were cultured on irradiated mouse embryonic fibroblasts (R and D Systems, Minneapolis, MN, USA) in miPSC medium: knockout DMEM with 4.5 g/ L d-glucose (Gibco, Grand Island, NY, USA), 10% knockout serum replacement (KSR) (Gibco), 10% fetal bovine serum (FBS) (HyClone, Logan, UT, USA), 1× MEM nonessential amino acids (MEM NEAA) (Gibco), 1× GlutaMAX (Gibco), 0.1 mM 2-mercaptoethanol (BME) (Life Technologies, Grand Island, NY, USA), and 0.02% ESGRO-LIF (Millipore, Billerica, MA, USA). Cells were incubated at 37 °C in 5% CO 2 .
3
Generating Mouse Induced Pluripotent Stem Cells
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We utilized mouse induced pluripotent stem cell lines derived at the UMN Stem Cell Institute as previously described (Terzic et al. 2016 (link)) from primary cultures of C57BL/6J and CAST/EiJ E13.5 mouse embryonic fibroblasts obtained from Jackson Laboratories (Bar Harbor, ME, USA), using octamer-binding transcription factor-4 (Oct4), sex-determining region y-box 2 (Sox2), and Kruppel-like factor-4 (Klf4) as reprogramming factors, introduced using pMXs retroviral vectors.
Induced pluripotent stem cells were cultured on irradiated mouse embryonic fibroblasts (R and D Systems, Minneapolis, MN, USA) in miPSC medium: knockout DMEM with 4.5 g/ L d-glucose (Gibco, Grand Island, NY, USA), 10% knockout serum replacement (KSR) (Gibco), 10% fetal bovine serum (FBS) (HyClone, Logan, UT, USA), 1x MEM nonessential amino acids (MEM NEAA) (Gibco), 1x GlutaMAX (Gibco), 0.1 mM 2-mercaptoethanol (BME) (Life Technologies, Grand Island, NY, USA), and 0.02% ESGRO-LIF (Millipore, Billerica, MA, USA). Cells were incubated at 37°C in 5% CO2.
Induced pluripotent stem cells were cultured on irradiated mouse embryonic fibroblasts (R and D Systems, Minneapolis, MN, USA) in miPSC medium: knockout DMEM with 4.5 g/ L d-glucose (Gibco, Grand Island, NY, USA), 10% knockout serum replacement (KSR) (Gibco), 10% fetal bovine serum (FBS) (HyClone, Logan, UT, USA), 1x MEM nonessential amino acids (MEM NEAA) (Gibco), 1x GlutaMAX (Gibco), 0.1 mM 2-mercaptoethanol (BME) (Life Technologies, Grand Island, NY, USA), and 0.02% ESGRO-LIF (Millipore, Billerica, MA, USA). Cells were incubated at 37°C in 5% CO2.
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