Hct116 and U2OS cell lines with a TetR repressor were kindly provided by Stephan Geley, SV40 MEF were generated by standard methods36 (link) and cultured in DMEM (Sigma) supplemented with 10% FCS (PAA), Penicillin/Streptomycin (Sigma) and 250 μM L-Gln (Invitrogen). To generated inducible shPDCD5 knockdown lines, shRNAs (human: shPDCD5-1: 5′- GCA GAA ATG AGA AAC AGT ATC -3′; shPDCD5-2 5′- CAG ATG GCA AGA TAT GGA CAA -3′; mouse: shPDCD5-1: 5′- GCA GAA ATG AGA AAC AGT ATC -3′; shPDCD5-2 5′- GAA CAA GGT TTG ATA GAA A -3′) were cloned into the lentiviral vector pHR-Dest-SFFV-puro40 (link) by Gateway cloning (Invitrogen). Stable cell lines were generated by lentiviral transduction of Hct116, U2OS and MEF cells followed by selection with puromycin. To induce PDCD5 knockdown, cells were treated with 1 μg/ml doxocyclin for 5 days before assays were performed. A549 cells were cultured in DMEM (Sigma) supplemented with 10% FCS (PAA), Penicillin/Streptomycin (Sigma) and 250 μM L-Gln (Invitrogen). Lymphoma cell lines PreB697 and CEM were cultured in RPMI 1640 Medium supplemented with 10% FCS (PAA), Penicillin/Streptomycin (Sigma) and 250 μM L-Gln (Invitrogen). PDCD5 overexpressing lines were generated by Lentiviral transduction with 3xFlag-PDCD541 (link).
L gln
Manufactured by Thermo Fisher Scientific
Sourced in United States
L-Gln is a laboratory reagent used in cell culture applications. It serves as a source of the amino acid glutamine, which is essential for cell growth and metabolism.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Streptomycin, by Thermo Fisher Scientific (3 mentions)
Penicillin, by Thermo Fisher Scientific (3 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions)
Trypsin edta, by Thermo Fisher Scientific (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Gemini Bio (2 mentions)
2 mercaptoethanol, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Stem cell factor (scf), by Thermo Fisher Scientific (2 mentions)
Interleukin 3 (il 3), by Thermo Fisher Scientific (2 mentions)
P s imdm, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Htb 38, by Thermo Fisher Scientific (1 mentions)
Hek293, by American Type Culture Collection (1 mentions)
Mcf 10a, by American Type Culture Collection (1 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (1 mentions)
22 protocols using l gln
1
Inducible PDCD5 Knockdown Cell Lines
2
Culturing Intestinal Epithelial Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Caco-2 cell line (ATCC® HTB-37 ™) isolated from a human colorectal adenocarcinoma was cultured in a Dulbecco’s Modified Medium F-12 Nutrient Mixture (DMEM F-12) supplemented with 10% Fetal Bovine Serum (FBS; Invitrogen, USA). Caco-2 cells exhibit spontaneous epithelial differentiation in vitro. Another human colorectal adenocarcinoma, the HT29 cell line (ATCC® HTB-38 ™), cultured in DMEM F-12 supplemented with 10% FBS and 1% L-glutamine (L-Gln; Invitrogen) was used as control in some experiments.
The Simian Vero-E6 renal epithelial cell line (ATCC® CRL-1586 ™), isolated from Chlorocebus sabaeus (African green monkey) was cultured in minimum essential medium (MEM. Gibco; Invitrogen) containing 4% FBS and 1% L-glutamine (L-Gln; Invitrogen). The HCT-8 (ATCC® CCL-244™), a human tumor epithelial cell line isolated from an ileal colorectal adenocarcinoma, was cultured in Roswell Park Memorial Institute 1640 medium (RPMI) (Gibco, Thermo Fischer) supplemented with 10% FBS.
All cells were cultured in a 175-cm2 flasks at 37°C in a 5% CO2 atmosphere. Every two days the medium was replenished, and confluent cultures were sub-cultured after harvesting of adherent cells by trypsination (0,05% Trypsin-EDTA, Invitrogen, USA).
The Simian Vero-E6 renal epithelial cell line (ATCC® CRL-1586 ™), isolated from Chlorocebus sabaeus (African green monkey) was cultured in minimum essential medium (MEM. Gibco; Invitrogen) containing 4% FBS and 1% L-glutamine (L-Gln; Invitrogen). The HCT-8 (ATCC® CCL-244™), a human tumor epithelial cell line isolated from an ileal colorectal adenocarcinoma, was cultured in Roswell Park Memorial Institute 1640 medium (RPMI) (Gibco, Thermo Fischer) supplemented with 10% FBS.
All cells were cultured in a 175-cm2 flasks at 37°C in a 5% CO2 atmosphere. Every two days the medium was replenished, and confluent cultures were sub-cultured after harvesting of adherent cells by trypsination (0,05% Trypsin-EDTA, Invitrogen, USA).
3
Cell Line Culture Conditions Comparison
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MCF-10A, HEK293 and SW480 cells were purchased from the ATCC. MDA-MB-468 was kindly donated by Dr. L. Planelles (CNB/CSIC) and Caco-2 by Dr. A. González (CNB/CSIC). SW480 cells were maintained in DMEM (Lonza) supplemented with 10% FBS (GBio) and 2 mM L-Gln (Gibco). Caco-2 cells were maintained with 1 mM sodium pyruvate (Gibco). MCF-10A cells were maintained in DMEM/F12 (Lonza, 1:1, v/v), 5% horse serum (Gibco), 2 mM L-Gln, insulin-transferrin-selenium (Gibco), 500 ng/ml hydrocortisone (Calbiochem) and 20 ng/mL epidermal growth factor (EGF, Upstate). MDA-MB-468 cells were cultured in Leibovitz's medium (L15, Lonza). All cell lines were maintained at 37ºC and 5% CO2 except MDA-MB-468, which was cultured without CO2 in L15 medium. Identity of the breast-derived cell lines was confirmed by STR genotyping (Genomics Facility, IIB/CSIC, Madrid, Spain).
4
Cell Culture Protocol for Various Cell Lines
5
Optimized CHO Cell Cultivation Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CHO-S (cGMP Banked, Cat. No. A13696-01), was purchased from Thermo Fisher Scientific (USA). CHO-K1 and S were maintained in CD-CHO medium (Thermo Fisher Scientific) supplemented with 8 mM L-Gln (Thermo Fisher Scientific, USA) and 0.2% Anti-Clumping Agent (Thermo Fisher Scientific, USA). Cells were grown in either TubeSpin Bioreactor 50 (TPP) tubes (TPP Techno Plastic Products, Switzerland) with a working volume of 15 mL or 125 mL shaking flasks (Corning, USA) with a working volume of 25–30 mL. The cell cultures in the tubes and shaking flasks were incubated at 37 °C, 7% CO2, and humidified air at a shaking speed of 250 and 140 rpm, respectively. The cells were passaged twice a week with a seeding density of 2 × 105 viable cells per mL. The cell cultures were mixed with trypan blue and counted by ViCELL XR Cell Counter (Beckman Coulter, Germany) for VCD and % viability measurement.
6
Culturing Diverse Melanoma Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SK-MEL-147, UACC-62, 1205-LU, 451-LU, WM164, WM793, HEK293T, GP2-293 and Phoenix-Ampho cells were grown in DMEM, high glucose, pyruvate, and GlutaMax (Gibco 31966021) supplemented with 10% heat-inactivated FBS (Gibco 10270106) and 1% penicillin-streptomycin (Gibco 15070063). Mel-ST cells were grown in the same conditions except for 7% FBS. Human cell lines were maintained at 37°C with 5% CO2 and were regularly tested for mycoplasma infection. Drosophila SL2 cells were grown in Schneider medium supplemented with L-Gln (ThermoFisher), 10% heat-inactivated FBS and 1% penicillin-streptomycin at 25ºC. Melanoma SK-MEL-147, UACC-62, 451-LU, WM164, WM793 and 1205-LU cells were provided by Maria S. Soengas (CNIO), while Mel-ST cells were a gift of Corine Bertolotto (University of Nice, INSERM). All melanoma cell lines in this study were authenticated using the GenePrint® 10 System in the Genomics Unit of Centro Nacional de Investigaciones Oncológicas (CNIO).
7
Cell Lines Authentication and Maintenance
8
Cell Line Authentication and Maintenance
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
TC32, TC71, and A673 Ewing sarcoma cells were the gift of Dr. T. Triche (The Saban Research Hospital, Children’s Hospital of Los Angeles, CA). EW8 Ewing sarcoma cells were the gift of Peter Houghton (Nationwide Children’s Hospital, Columbus, OH). The MCF7 breast carcinoma cell line was the gift of Dr. Patricia Steeg (National Cancer Institute, Bethesda, MD). The RH30 and RD rhabdomyosarcoma cells were the gift of Lee Helman (National Cancer Institute, Bethesda, MD). The A2058 melanoma cell line was obtained from the American Type Culture Collection (ATCC, Manassas, VA). The identity of all cells was independently authenticated by short tandem repeat genotyping. All cells were maintained in culture in RPMI 1640 (Invitrogen, Carlsbad, CA). The medium was supplemented with 10% fetal bovine serum (Gemini Bio-Products, West Sacramento, CA), 2 mM L-Gln, 100 U/mL of penicillin and 100 μg/mL of streptomycin (ThermoFisher, Waltham, MA).
9
Huh-7.5 Cell Line Maintenance
10
Fatty Acids Impact on Iron Metabolism
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human HepG2 hepatocytes were grown in DMEM medium supplemented with 10% fetal calf serum (FCS), 1% Penicillin Streptomycin and 1% L-GLN (Life Technologies) in a humidified atmosphere with 5% CO2 at 37°C. Viability was assessed by Trypan blue exclusion dye test. In order to evaluate the effect of increased exposure to fatty acids on iron metabolism, cells were treated with palmitic acid (PA 0.66 mM: saturated, C16:0) plus oleic acid (OA 1 mM: n-9 mono-unsaturated, C18:1, cis) 24 hours before the experiments. To assess the effect of specific types of fatty acids on iron metabolism, HepG2 cells were also treated for 24 hours with the following: linolenic acid (LnA 0.025 mM; n-3 unsaturated, 18:2), stearic acid (SA 0.025 mM: saturated, C18:0), elaidic acid (EA; 0.1 mM: n-9 mono-unsaturated, 18:1, trans), lauric acid (LA 0.2 mM: saturated medium-chain, C12:0). Iron was supplemented for 24 hours as ferric ammonium citrate (FAC) 150 μM [27 (link)]. Intracellular iron concentration was measured by atomic absorption spectrometry [25 (link)]. For lipid visualization and quantification through Oil Red O staining, after being washed 3x with PBS, cells were fixed with 10% formalin, stained with 0.7% Oil Red O solution (Sigma, St. Louis, MO) and absorbance was then read at 540 nm.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!