U87 (ATCC HTB-14) and T98G (ATCC CRL-1690) cells were obtained from the American Type Culture Collection (ATCC, Manassas, USA) and genotyped (Genolytic GmbH, Leipzig, Germany) to confirm their identity prior to the experiments. Cells were propagated in 250 ml culture flasks (Sarstedt AG & Co., Nümbrecht, Germany) using 10 ml of standard culture medium (SCM) consisting of DMEM (Dulbecco’s Modified Eagle Medium) with 4.5 g/l glucose, and without pyruvate (Life Technologies, Darmstadt, Germany), supplemented with 10% fetal bovine serum (FBS superior, Biochrom, Berlin, Germany), 2 mM GlutaMAX (Life Technologies) and Penicillin-Streptomycin (Life Technologies) at 37°C and 5% CO2 in humidified air in an incubator.
250 ml culture flasks
Manufactured by Sarstedt
Sourced in Germany
250 ml culture flasks are laboratory equipment used for the cultivation and growth of cells, tissues, or microorganisms. They provide a contained environment with a defined volume for culturing purposes.
Automatically generated - may contain errors
Lab products found in correlation
Fbs superior, by Harvard Bioscience (4 mentions)
Glutamax, by Thermo Fisher Scientific (4 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Pyruvate, by Thermo Fisher Scientific (1 mentions)
T98g atcc crl 1690, by American Type Culture Collection (1 mentions)
U87 atcc htb 14, by American Type Culture Collection (1 mentions)
Powerplex 21 system, by Promega (1 mentions)
4 protocols using 250 ml culture flasks
1
Cell Line Authentication and Culturing
2
Cell Culture of Glioblastoma Cell Lines
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U87 and T98G cells were obtained from the ATCC (Manassas, VA, USA) and the line LN405 was obtained from the DMSZ (Braunschweig, Germany). All cell lines were genotyped (Genolytic GmbH, Leipzig, Germany) to confirm their identity. Cells were propagated in 250 mL culture flasks (Sarstedt AG & Co., Nümbrecht, Germany) using 10 mL of DMEM containing 4.5 g/L glucose, without pyruvate (Life Technologies, Darmstadt, Germany), supplemented with 10% fetal bovine serum (FBS superior, Biochrom, Berlin, Germany), 2 mM GlutaMAX (Life Technologies) and Penicillin-Streptomycin (Life Technologies) at 37 °C and 5% CO2 in humidified air in an incubator.
3
Glioblastoma Cell Culture Protocol
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All cells were propagated in 250 ml culture flasks (Sarstedt AG & Co., Nümbrecht, Germany) using 10 ml of standard culture medium (SCM: DMEM / 4.5 g/l glucose, without pyruvate (Life Technologies, Darmstadt, Germany) supplemented with 10% fetal bovine serum (FBS superior, Biochrom, Berlin, Germany), 2 mM GlutaMax (Life Technologies) and Penicillin-Streptomycin (Life Technologies)) at 37°C and 5% CO2 in humidified air in an incubator. The human glioblastoma multiforme (GBM) cell lines T98G and U87 were obtained from ATCC (Manassas, USA) and the line LN405 from the German collection of microorganisms and cell cultures (DSMZ, Braunschweig, Germany). All cells were genotyped (Genolytic GmbH, Leipzig, Germany) and their identity confirmed.
4
Starvation-Induced Metabolic Shifts in U87 Cells
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U87 cells were originally obtained from the ATCC (Manassas, USA) and cultured in 250 mL culture flasks (Sarstedt AG & Co., Nümbrecht, Germany) using DMEM/25 mM glucose, without pyruvate (Life Technologies, Darmstadt, Germany) supplemented with 10 % fetal bovine serum (FBS superior, Biochrom, Berlin, Germany), 2 mM GlutaMAX and antibiotics (Life Technologies) at 37 °C and 5 % CO2 in humidified air in an incubator. In order to confirm identity over long culture periods, cells were genotyped by STR analysis at the Genolytic GmbH (Leipzig, Germany) using a PowerPlex® 21 System (Promega, Mannheim, Germany) and cells were confirmed as the U87MG cell line from the ATCC [17 (link)]. For starvation experiments, U87 cells were seeded in 6-well plates (TPP, Trasadigen, Switzerland) at a density of 106 cells per well in 2 mL full supplemented DMEM (10 % FBS, GlutaMAX, antibiotics) and incubated for 3 h before receiving fresh medium (1 mL) without a carbon source and without GlutaMAX and FBS (DMEM [0]). Cells were incubated for 20 h before replacing the culture medium with medium (1 mL) containing either different concentrations of glucose or medium without glucose but pyruvate at a concentration of 5 mM.
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