HepG2 cells were cultured in DMEM (GIBCO by Life Technologies) with 10% heat-inactivated FCS (ThermoFisher Scientific). BECs were cultured in Hams F12 Nutrient Mixture (GIBCO by Life Technologies) with the following supplements: 10% heat-inactivated human serum (TCS Biosciences), hepatocyte growth factor (HGF; 10 ng/ml; Peprotech), epidermal growth factor (EGF; 10 ng/ml; Peprotech), hydrocortisone (2 µg/ml; Sigma-Aldrich), cholera toxin (10 ng/ml; Sigma-Aldrich), tri-iodo-thyronine (2 nM; Sigma-Aldrich), insulin (0.124 U/ml; Sigma-Aldrich), 100 U/ml penicillin, 100 μg/ml streptomycin, 2 mM L-glutamine [penicillin-streptomycin-glutamine (100×); GIBCO by Life Technologies]. HSECs were maintained in human endothelial serum-free media (SFM) with 10% human serum, 100 U/ml penicillin, 100 μg/ml streptomycin, 2 mM L-glutamine (all GIBCO by Life Technologies), 10 ng/ml of HGF (Peprotech), and 10 ng/ml VEGF (Peprotech). HSCs and aLMFs were cultured in Dulbecco's Modified Eagle medium (DMEM; GIBCO by Life Technologies) with 16% heat-inactivated FCS (ThermoFisher Scientific) and 100 U/ml penicillin, 100 μg/ml streptomycin, and 2 mM L-glutamine (all GIBCO by Life Technologies).
Penicillin streptomycin glutamine 100
Manufactured by Thermo Fisher Scientific
Sourced in United States
Penicillin-streptomycin-glutamine (100×) is a ready-to-use solution that contains a combination of antibiotics and L-glutamine. It is commonly used as a supplementary component in cell culture media to provide antimicrobial protection and support cellular growth.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (2 mentions)
Dulbecco s modified eagle medium dmem, by Thermo Fisher Scientific (2 mentions)
Ham s f 12 nutrient mixture, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Cholera toxin, by Merck Group (1 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions)
Vascular endothelial growth factor (vegf), by Thermo Fisher Scientific (1 mentions)
Hepatocyte growth factor (hgf), by Thermo Fisher Scientific (1 mentions)
Insulin, by Merck Group (1 mentions)
Hydrocortisone, by Merck Group (1 mentions)
Triiodothyronine, by Merck Group (1 mentions)
Etoposide, by Enzo Life Sciences (1 mentions)
Protein a g column, by Thermo Fisher Scientific (1 mentions)
Protein l column, by Thermo Fisher Scientific (1 mentions)
Hek293t 17 cell line, by American Type Culture Collection (1 mentions)
Bicinchoninic acid bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Steady glo luciferase reagent, by Promega (1 mentions)
14 protocols using penicillin streptomycin glutamine 100
1
Culturing Primary Liver Cell Types
2
Ruthenium-labeled Protein A/G/L Reagent
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Unlabeled protein A/G/L used for detection in the NHP TAb assay was purchased from BioVision (Milpitas, CA, USA). The ruthenium-labeled protein A/G/L reagent was prepared by reconstituting protein A/G/L according to manufacturer’s recommendation at 10 mg/ml in DPBS without calcium and magnesium. The concentration was adjusted to 1 mg ml−1 in DPBS and labeled with 10 nmol MSD Sulfo-NHS Ruthenium for 1 h at ambient temperature with rotation. The labeled protein A/G/L was purified by three cycles of filtration with Amicon-15 centrifugal filters using DPBS and stored at −80 °C in single-use aliquots. Tris Buffered Saline with 1% (w/v) Casein (TBS-C) was from BioRad (Hercules, CA, USA). The protein A/G columns, protein L columns, and bicinchoninic acid (BCA) protein assay kits were from Thermo Fisher Scientific (South San Francisco, CA, USA). Etoposide was from Enzo Life Sciences (Farmingdale, NY, USA), Steady-Glo Luciferase reagent was from Promega (Sunnyvale, CA, USA), fetal bovine serum (FBS) without heat inactivation was from HyClone (Logan, UT, USA), and Penicillin-Streptomycin-Glutamine (100 ×) was from Gibco (South San Francisco, CA, USA). The HEK293T/17 cell line was purchased from ATCC (Manassas, VA, USA). The GFP quantification kit was purchased from Cell Biolabs (San Diego, CA, USA).
3
Cell Culture Conditions and Sources
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A549, ZR-75-1, MDA-MB-231, and MCF-7 cells were obtained from the Bioresource Collection and Research Centre (BCRC) in Hsinchu, Taiwan. The paclitaxel-resistant 7TR cells were generated by treating MCF-7 with increased concentrations (0, 5, 10, 20, 40, 60, 80, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280, and 300 nM) of paclitaxel continuously and selecting the survived cells. HEK-293T was a gift from Dr. Shih-Hsiung Wu (Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan). The human lung adenocarcinoma cell lines with less invasive (CL1-0) and highly invasive capacities (CL1-5) were established previously [56 (link)]. A549 cells were cultivated in F-12K, MCF-7 and 7TR in MEM, ZR-75-1, CL1-0, and CL1-5 in RPMI-1640, MDA-MB231 and HEK-293T in DMEM. Each medium was supplemented with 10% fetal bovine serum (FBS) that was heat-inactivated at 56 °C for 30 min in a water bath and 1% of a commercial product [penicillin-streptomycin-Glutamine (100×)] (Gibco catalog number: 10378016). All cell culture media and supplements were purchased from Gibco (Carlsbad, CA, USA). The cells were maintained in the presence of 5% CO2 at 37 °C.
4
Culturing Breast Cancer Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MCF-7 (provided by American Type Culture Collection, ATCC, Manassas, Virginia, USA), MDA-MB-436 (provided by University Clinic Ulm, Ulm, Germany), MDA-MB-453 (provided by University Clinic Ulm, Ulm, Germany), MDA-MB-468 (provided by University Clinic Ulm, Ulm, Germany), and ZR75-1 (provided by Experimental Pharmacology and Oncology, Berlin-Buch, Berlin, Germany) were cultured in DMEM with L-glutamine (Gibco/ThermoFisher Scientific, Waltham, MA, USA), 10% FBS (Pan Biotech, Aidenbach, Germany), 1.2% L-glutamine (Gibco/ThermoFisher Scientific), 1.0% Penicillin-Streptomycin-Glutamine (100×) (Gibco/ThermoFisher Scientific), 1.0% MEM NEAA (non essential amino acid) (Gibco/ThermoFisher Scientific), 0.1% human recombinant insulin (Gibco/ThermoFisher Scientific), and 0.1% hEGF (Sigma-Aldrich/Merck, St. Louis, MO, USA) (Table 1 ) [22 (link),23 (link)]. HCC-1937 cells (provided by ATCC) were cultured in RPMI 1640 medium with 15% FBS (Pan Biotech, Aidenbach, Germany) and 1% of Penicillin-Streptomycin-Glutamine (100X) (Gibco/ThermoFisher Scientific). Cells were cultured in a humid 5% CO2 incubator at 37 °C and all cell lines were negative for mycoplasma, verified by PCR.
5
Laparoscopic Ovarian Biopsy for HGSOC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Laparoscopic ovarian biopsies, before and after chemotherapy, were obtained from a cohort of five EOC patients, classified as HGSOC (Table 1 ). All five patients involved in this study gave their written consent. The study design was approved by the Ethics Committee of Policlinico Umberto I Hospital, C.E. Ref: 1454/24.07.08, Prot. no. 702/08 (Rome, Italy). Biopsies were snap-frozen and stored at −80 °C. To identify differences in gene expression by chemotherapy, we used biopsies from patients before chemotherapy treatment as controls. After de-bulking surgery and biopsies, tumor cells were classified as Grade 3, known also as high grade ovarian cancer cells. SKOV3 is a wtBRCA1 gene-carrying ovarian serous adenocarcinoma cell line and was purchased from ATCC® (HTB-77™). With one exception, the patients did not present with BRCA1/2 mutations. The cells were cultured in RPMI-1640, (Sigma-Aldrich; Merck Life Science S.r.l., Milan, Italy, Cat. n.R0883), supplemented with 10% FBS (Corning™; Cat.n.15377636), and 100 U/mL penicillin, 100 µg/mL streptomycin, and 29.2 mg/mL of l -glutamine (Gibco™ Penicillin-Streptomycin-Glutamine (100×) Cat.n.10378-016).
6
Culture of Human Retinoblastoma Y79 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human retinoblastoma cell line Y79 was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). The cells were cultured in RPMI-1640 medium containing 10% (v/v) fetal bovine serum (FBS) (GIBCO BRL, Grand Island, NY, USA) and 1% (v/v) penicillin-streptomycin-glutamine (100×) (GIBCO BRL). The cell culture was performed in an incubator at 37°C with 5% CO2 (Thermo Fisher Scientific, Waltham, MA, USA).
7
Cell Culture Media and Reagents
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Dulbecco modified Eagle’s medium (DMEM), Ham F12-K medium, Rosselle’s Park Memorial Institute medium (RPMI-1640) and penicillin/streptomycin/glutamine (100×) were purchased from GIBCO BRL (UK). Fetal bovine serum (FBS) and trypsin/EDTA were purchased from Hyclone (USA) and BDH Chemicals (UK), respectively. Dimethyl sulphoxide (DMSO), hydrogen peroxide (H2O2) and tamoxifen were purchased from Sigma-Aldrich (USA). Phosphate buffered saline (PBS) was purchased from Amresco (USA). All chemicals used in the experiments were of analytical grade.
8
Exosome Isolation and Characterization Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DMEM and penicillin-streptomycin-glutamine (100×) were purchased from Gibco (Waltham, MA, USA). Fetal bovine serum (FBS) was purchased from Procell Life Science & Technology Co.,Ltd (Wuhan, China). Exosome-depleted FBS was purchased from Cellmax Cell Technology Co., Ltd. (Beijing, China). Lefty1 recombinant plasmid (Gene ID: 498299), negative plasmid and GAPDH primers, U6 primers, α-SMA primers, lefty1 primers were purchased from Genepharma Technology Co., Ltd (Shanghai, China). PKH26 was got from Sigma (St. Louis, MO, USA). TransExo™ serum/plasma exosome total RNA extraction kit and PrimeScript RT master mix were got from TransGen Biotech Co., Ltd (Beijing, China). Plasmocin™ prophylactic, dextran-AlexaFluor555, transferrin-AlexaFluor 555, cholera toxin subunit B-AlexaFluor 555 and DiR were purchased from Invivogen (San Diego, CA, USA). Hoechst33342 and lysotracker green were purchased from Beyotime Biotechnology Co., Ltd. (Shanghai, China). Anti-CD9(ab92726), CD81(ab109201), TSG101 (ab125011), α-SMA(ab7817), collagen I(ab270993) and HRP coupling goat anti-rabbit IgG (ab205718), goat anti-rabbit IgG H&L (Alexa Fluor 488) (ab150077) were purchased from Abcam (Cambridge, UK). Anti-alix (T57215) was purchased from Abmart Medical Technology Co., Ltd. (Shanghai, China). Anti-CD63(AF5117), calnexin (AF5362) were purchased from Affinity Biosciences, Inc. (Cincinnati, OH, USA).
9
Establishment of Thyroid Cancer Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human thyroid cancer cell line BHP10-3 was gained from Qilu Hospital and IHH4 was purchased from American Type Culture Collection. All cells were maintained at 37 °C, and 5% CO2 in RPMI-1640 (C11875500BT, GIBCO) supplemented with 10% fetal bovine serum (FBS) (10270106, GIBCO) and penicillin-streptomycin-glutamine (100×) (10378016, GIBCO). BHP10-3 cell lines with stable knockdown of SHMT2 (LV-shSHMT2) and stable overexpression of SHMT2 (LV-SHMT2) and control cell lines were established and preserved in our laboratory. Briefly, lentiviruses infected cell pools were selected using 1 μg/mL puromycin (S7417, Selleck) for 2 weeks. The overexpressed and knockdown efficiency was probed by RT-qPCR or western blot analysis.
10
Isolation and Culture of Human USSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human USSCs were isolated from normal human cord blood provided by the obstetrics and gynaecology ward, Shariati hospital, Tehran‐Iran. Cord blood was collected from the mother and informed written consent was obtained. The mononuclear cells were obtained by Ficoll gradient separation (Biochrom) followed by RBCs lysis using ammonium chloride. Cells were cultured in T25 culture flasks (Costar) at a confluency of 5–7 × 106 cells/ml. Cells were then cultured in myelocult medium (StemCell Technologies), low glucose DMEM, Glutamax (Thermo Fischer), Penicillin–Streptomycin‐Glutamine (100×, Thermo Fischer) and incubated at 37°C in 5% CO2 in a humidified atmosphere. Cultured cells were later detached when they reached 80% confluency using 0.25% trypsin.
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!