Macrophages were obtained as described previously (4 (link)). TGC peritoneal macrophages (pMø) were obtained from mice after intraperitoneal (i.p.) injection with 4% Difco™ Fluid TGC Medium [Becton, Dickinson and Company (BD), Franklin Lakes, NJ, USA], and maintained in HyClone RPMI 1640 medium (GE Healthcare, Chicago, IL, USA), supplemented with 0.5% fetal bovine serum (FBS), 2 mM L-glutamine, 5 mM HEPES, and 1 mM sodium pyruvate (all from Thermo Fisher Scientific, Waltham, MA, USA) (complete medium) in a 95% air–5% CO2 incubator at 37°C. For further selection, the cell suspension was incubated to allow pMø to adhere to the culture dish and then proceed with the analyses. RAW264.7 cells were maintained in DMEM (GIBCO, Thermo Fisher Scientific) with 10% FBS at 37°C in a humidified incubator containing 5% CO2. For stimulation, 1 or 0.5 × 106 RAW264.7 cells or pMø, in complete medium with 0.5% FBS per well, were activated with the indicated stimuli and time points.
Hyclone rpmi 1640 medium
Manufactured by GE Healthcare
Sourced in United States, China
HyClone™ RPMI-1640 medium is a cell culture medium commonly used for the in vitro cultivation of a variety of cell types, including lymphocytes and other suspension cells. It provides a standardized and defined nutritional environment to support cell growth and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (18 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (7 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions)
Penicillin, by Thermo Fisher Scientific (4 mentions)
Streptomycin, by Thermo Fisher Scientific (4 mentions)
L glutamine, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by GE Healthcare (2 mentions)
Gentamicin, by Merck Group (2 mentions)
Penicillin, by GE Healthcare (2 mentions)
Streptomycin, by GE Healthcare (2 mentions)
Mda mb 231, by American Type Culture Collection (2 mentions)
Mcf 7, by American Type Culture Collection (2 mentions)
Sw480, by American Type Culture Collection (2 mentions)
Sw620, by American Type Culture Collection (2 mentions)
Hyclone, by GE Healthcare (2 mentions)
Hepes, by Thermo Fisher Scientific (1 mentions)
Foetal calf serum, by Merck Group (1 mentions)
Foetal calf serum, by Thermo Fisher Scientific (1 mentions)
Valproic acid, by Merck Group (1 mentions)
Rpmi 1640 1x medium, by Thermo Fisher Scientific (1 mentions)
38 protocols using hyclone rpmi 1640 medium
1
Isolation and Activation of Macrophages
2
Acute myeloid leukemia cell culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
NB4 human APL cells were a gift from Dr. David Scheinberg (Memorial Sloan Kettering Institute, New York, NY, USA). The ATRA-resistant NB4R cell line was a gift from Prof. Pier Paolo-Pandolfi (Beth Israel Deaconess Cancer Centre, Boston, MA, USA). Cells were cultured at 37°C, 5% CO2 in HyClone RPMI 1640 medium (GE Healthcare Life Sciences SH30027), supplemented with 10% foetal calf serum (Sigma-Aldrich, F7524) and 1% penicillin/streptomycin (Invitrogen, 10378-016). THP-1 cells (monocytic-lineage) were from the American Type Culture Collection (ATCC). Cells were cultured in RPMI 1640 (1x) medium (Gibco by Life technologies, 21870-076), supplemented with 10% foetal calf serum and 1% penicillin/streptomycin. Cells were seeded at 5x104 cells/ml prior to treatment. ATRA (Sigma-Aldrich, R2625) was used to induce differentiation at 1 μM, diluted from a 1 mM stock in 100% ethanol (EtOH). Control populations were treated with 0.1% v/v EtOH. Valproic acid (Sigma-Aldrich, P4543) was used at 1 mM concentration, diluted from a 500mM stock in H20.
3
Culturing Human Ovarian Cancer Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human ovarian cancer cell lines were obtained from the American Type Culture Collection (ATCC) and the University of Texas MD Anderson Cancer Center Characterized Cell Line Core Facility. Cell lines were routinely identified via short tandem repeat DNA profiling carried out by the Characterized Cell Line Core Facility at MD Anderson. Primary FTE cells were a gift from J. Liu from the Department of Pathology at MD Anderson. For all cell lines, mycoplasma testing was done using the ATCC PCR Universal Mycoplasma Detection Kit (30-1012K). OVCAR-8 cells were cultured in HyClone RPMI 1640 medium (SH30027.01, GE Healthcare Life Sciences) supplemented with 15% fetal bovine serum (FBS) (Sigma-Aldrich) and 0.2% gentamicin (50146970, Thermo Fisher Scientific). OVCAR-5 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; 10-013-CV; Corning) supplemented with 10% FBS and 0.2% gentamicin. FTE cells were cultured in medium 199 with MCDB 105 (1:1) with 10% FBS and 0.2% gentamicin. All cells were grown in humidified incubators kept at 37°C with 5% CO2.
4
Culturing Human Pancreatic Cancer Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human PDAC cell lines AsPC-1, BxPC-3, Capan-1, MIA PaCa-2, PCT-3, and SU.86.86 were donated by the University of Heidelberg. The cells were cultured in HyClone RPMI-1640 medium (GE Healthcare Life Sciences, Logan, Utah, USA) supplemented with 10% HyClone fetal bovine serum (FBS, GE Healthcare Life Sciences, Logan, Utah, USA) in a humidified incubator with 5% CO2 at 37 °C.
5
Esophageal Cell Line Culture Protocols
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
ESCC cell lines KYSE150, KYSE180, KYSE450, and TE-1 and immortal embryonic esophageal epithelium cell lines NE3 and HET-1A used in the research were purchased from the Cell Bank of Type Culture Collection of the Chinese Academy of Sciences. The cell line 293T was from our lab. The cell lines KYSE150, KYSE180, KYSE450, and TE-1 were cultured in HyClone™ RPMI-1640 medium, and the cell lines HET-1A and 293T were cultured in HyClone™ DMEM/High Glucose medium (GE Healthcare Life Sciences, HyClone Laboratories, Logan, UT, USA). The culture was with 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA). The cell line NE3 was cultured in a 1:1 mixture of EpiLife medium (Cascade Biologics, Inc., Portland, OR, USA) and defined keratinocyte serum-free medium (dKSFM; Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA). All of the cell lines were cultured at 37°C, 5% CO2. The culture medium was replaced according to the cell state. Subculture was carried out when the cell fusion was about 80%–90%.
6
Cryopreservation and Thawing of Cell Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Some samples were frozen for later analysis, all others were characterized fresh after processing. Cells were cryopreserved in HyClone RPMI-1640 medium (GE Healthcare Life Sciences) containing 10% heat-inactivated AB serum (Karolinska University Hospital, Huddinge, Sweden) and 10% CryoSure dimethyl sulfoxide (WAK-Chemie Medical GmbH). Contents were mixed and kept overnight in −80° C and thereafter kept in −192° C until use. Cells were thawed at RT in AB serum-supplemented RPMI-1640 medium and washed twice in PBS before extracellular staining.
7
ESCC Cell Line Culture Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All ESCC cell lines used in the study were obtained from the Translational Medicine Research Center, Shanxi Medical University (Taiyuan, China) and cultured in HyClone™ RPMI-1640 medium (GE Healthcare Life Sciences, HyClone Laboratories, Logan, UT, USA) with 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) at 37°C in a 5% CO2 incubator. Culture medium was replaced every two to three days. Subculture was carried out when the cells were fused to 80–90% confluency and logarithmic phase cells were used in the following experiments.
8
CLL Patient B-cell Isolation and Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Patient samples were obtained from Dept. of Internal Medicine–Hematology and Oncology, University Hospital Brno. B-cells were isolated from the peripheral blood of CLL patients undergoing monitoring and treatment at the hospital, as described here (Kaucká et al., 2013 (link)). CLL samples were obtained after written informed consent in accordance to the Declaration of Helsinki and by following protocols approved by the ethical committee of the University Hospital, Brno. Primary CLL cells were grown in HyClone™ RPMI 1640 medium (GE Healthcare) supplemented with 10% heat-inactivated FBS and 1% Penicillin/Streptomycin at 37°C and 5% CO2. Patient characteristics are available in the Supplementary Material .
9
CD34+ HSPC Culture and Genetic Manipulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
K562 cells, kindly provided by Prof. Ewa Sitnicka Quinn, were maintained in HyClone RPMI-1640 Medium (GE Healthcare Life Sciences) supplemented with 10% heat-inactivated FBS (GE Healthcare Life Sciences) and 1% Penicillin–Streptomycin (GE Healthcare Life Sciences). CD34+ HSPCs were cultured in Serum-Free Expansion Medium (SFEM, STEMCELL Technologies) supplemented with 1% Penicillin–Streptomycin (GE Healthcare Life Sciences), stem cell factor (SCF), FLT3-ligand (FLT3L) and thrombopoietin (TPO). All cytokines were used in the concentration 100 ng/ml and acquired from Peprotech. CD34+ cells were thawed and pre-stimulated in culture medium at 37 °C, 5% CO2 for 48 h prior to RNP electroporation and for 24 or 48 h prior to lentiviral sgRNA transduction.
10
RB Cell Line Culture and Reagent Preparation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Que (>99% pure) was purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany), dissolved in dimethyl sulfoxide (DMSO), aliquoted and stored at −22°C. Antibodies against VEGFR (cat. no. ab36844) and GAPDH (cat. no. ab8245) were purchased from Abcam (Cambridge, UK). The human RB Y79 cell line (Shanghai Bioleaf Biotech Co., Ltd., Shanghai, China), preserved at the Second Hospital of Shandong University (Jinan, China), was cultured in HyClone™ RPMI-1640 medium (GE Healthcare, Logan, UT, USA) supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) and 1% (v/v) antibiotics mixture (100 U/ml penicillin and 100 U/ml streptomycin) in an atmosphere of 95% air and 5% CO2 at 37°C.
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!