Vero (African green monkey kidney) cells were grown in Opti-Pro medium (Invitrogen) supplemented with 50 µg/ml of gentamicin as previously described (39 (link)). For ZIKV infection, Vero cell medium was changed to complete Dulbecco's modified Eagle medium (DMEM [Invitrogen] supplemented with 10% fetal bovine serum [FBS] and 1× penicillin-streptomycin-glutamine solution [Invitrogen]). Mosquito-derived C6/36 cells (Aedes albopictus) and 15P-1 cells (mouse testis Sertoli cells; ATCC) were maintained in complete DMEM at 32°C and 5% CO2. Primary human foreskin fibroblasts (ATCC) and SH-SY5Y cells (human neuroblastoma, CRL-2266; ATCC) were maintained in complete DMEM at 37°C and 5% CO2. HTR-8/SVneo human trophoblasts (CRL3271; ATCC) and JAR human placenta cells (HTB-144; ATCC) were maintained in RPMI 1640 medium (ATCC) supplemented with 5% FBS and 1× penicillin-streptomycin solution (Invitrogen) at 37°C and 5% CO2. BeWo human placenta cells (CCL-98; ATCC) were maintained in Ham’s F-12K medium (ATCC) supplemented with 10% FBS and 1× penicillin-streptomycin-glutamine solution (Invitrogen) at 37°C and 5% CO2. JEG-3 human placenta cells (HTB-36; ATCC) were maintained in Eagle minimum essential medium (EMEM; ATCC) supplemented with 10% FBS and 1× penicillin-streptomycin solution at 37°C and 5% CO2.
Penicillin streptomycin glutamine solution
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom, United States
Penicillin-streptomycin-glutamine solution is a sterile, ready-to-use liquid that contains a combination of penicillin, streptomycin, and L-glutamine. It is commonly used as a supplement in cell culture media to provide antimicrobial protection and support cell growth and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (6 mentions)
Opti mem, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Merck Group (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Rpmi 1640 medium, by American Type Culture Collection (2 mentions)
Melphalan, by Merck Group (2 mentions)
Dld 1, by American Type Culture Collection (1 mentions)
Foetal calf serum, by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Euroclone (1 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (1 mentions)
Knockout serum replacement (ksr), by Thermo Fisher Scientific (1 mentions)
Human tgf β1, by R&D Systems (1 mentions)
Salmonella enterica serotype typhimurium, by Merck Group (1 mentions)
Escherichia coli serotype 055 b5, by Merck Group (1 mentions)
Tgf β1, by Merck Group (1 mentions)
Gentamycin, by Quality Biological (1 mentions)
27 protocols using penicillin streptomycin glutamine solution
1
Zika Virus Cell Culture Protocols
2
Isolation and Expansion of Primary Human Trabecular Meshwork Cells
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Primary HTM cells were cultured from TM tissue isolated from the leftover donor corneal rings after they had been used for corneal transplantation at the Indiana University Clinical Service, Indianapolis, as described previously (Keller et al., 2018 (link)). HIPPA compliance guidelines were adhered for the use of human tissues. The usage of donor tissues was exempt from the DHHS regulation and the IRB protocol (1911117637), approved by the Indiana University School of Medicine IRB review board. The age, race, and sex of the donors were obtained from eye banks, which provided the corneas. TM tissue extracted from the corneal ring was chopped into fine pieces and placed in a 2% gelatin‐coated six‐well tissue culture plate sandwiched by a coverslip. The tissues were grown in OptiMEM (Gibco, #31985‐070), containing 20% fetal bovine serum (FBS) and penicillin‐streptomycin‐glutamine solution (Gibco, #10378‐016). The expanded population of HTM cells was sub‐cultured after 1–2 weeks in Dulbecco's modified Eagle's medium, containing 10% FBS and characterized by detection of dexamethasone‐induced myocilin. All experiments in this manuscript were performed using biological replicates. The normal TM (NTM5) cell line obtained from Dr. Weiming Mao, Indiana University School of Medicine, was cultured as mentioned above.
3
Culturing CRC Cell Lines DLD-1 and HT-29
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Two CRC cell lines, namely DLD-1 and HT-29, were obtained from American Type Culture Collection. The cells were cultured in Dulbecco’s modified Eagle’s medium (Gibco, Thermo Fisher Scientific, UK), supplemented with 10% foetal calf serum (Merck, UK), and penicillin/ streptomycin/glutamine solution at 100 units ml−1, 100 μg ml−1 and 0.29 mg ml−1, respectively (Gibco, Thermo Fisher Scientific, UK) and were incubated at 37 °C in 5% CO2.
4
Generation and Characterization of SBDS Cell Lines
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HEK-293T and HeLa cells from ATCC were cultured in DMEM (Euroclone) supplemented with 10% fetal bovine serum (FBS) and penicillin/streptomycin/glutamine solution (GIBCO) at 37 °C under 5% CO2. Mycoplasma testing was performed before experiments. 293T cells were transfected at 60–70% confluence with pGIPZ eIF6 shRNA and pGIPZ scramble lentiviral vectors (second generation lentivirus from Open Biosystem) and with pFCY, SBDS shRNA and pFCY scramble lentiviral vector (a generous gift from the lab of DC. Link, Washington University School of Medicine) to produce lentiviral particles. HeLa cells were infected with lentivirus at a confluence of 50%. Experiments were performed after one week from infection. Silencing of protein was measured by Western Blot analysis.
SBDS cell lines were generated in our lab (Calamita P. et al., in preparation) by immortalizing MEFs with DNp53Ras retroviral vectors. The mutated line express in homozygosity the SBDS protein with the R126T point mutation. Primary MEFs belong to the SBDS mouse model of the laboratory of Joahanna Rommens (The Hospital of Sick Children, Toronto).
SBDS cell lines were generated in our lab (Calamita P. et al., in preparation) by immortalizing MEFs with DNp53Ras retroviral vectors. The mutated line express in homozygosity the SBDS protein with the R126T point mutation. Primary MEFs belong to the SBDS mouse model of the laboratory of Joahanna Rommens (The Hospital of Sick Children, Toronto).
5
Testis Organ Culture Protocol
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The organ cultures of the testis fragments used in this study were created according to a previously described method [31 (link)] with a few modifications. The culture medium was 10% KnockOut serum replacement (KSR; Gibco)/ alpha-Minimum Essential Medium (MEM; Gibco) supplemented with a MEM non-essential amino acid solution (x1, Gibco), 1mM sodium pyruvate (Gibco) and a penicillin/streptomycin/glutamine solution (x1, Gibco). The fragmented testis tissues were positioned on agarose blocks that were in a 12-well culture plate with the culture medium. The agarose blocks were constructed as follows; 1.5% (W/V) Agarose LO3 (Takara-Shuzo) was solidified in a 48-well culture plate, and the cylindrical agarose gels were pulled out and pre-equilibrated in the culture medium for more than 24 hrs.
6
Bone Marrow-Derived Dendritic Cell Differentiation and Activation
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Bone-marrow cells were cultured for 6 days in RPMI-1640 supplemented with 10% FBS (Hyclone), 1X Penicillin Streptomycin Glutamine solution (Gibco), and 1∶3000 GM-CSF supernatant (U-M Hybridoma core facility). GM-CSF-supplemented media was changed every second day (days 0, 2, 4, and 6). When indicated, 1–25 ng/ml human TGF-β1 (R&D Systems) was added on days 0, 2, 4, and/or 6. On day 7, bone-marrow-derived DCs were analyzed by flow cytometry. To examine DC activation, bone-marrow-derived DCs were stimulated on day 6 with 1 or 100 ng/ml LPS from Escherichia coli (serotype 055:B5) (Sigma) or Salmonella enterica (serotype typhimurium) (Sigma) or 2∶1 ratio LPS-infected apoptotic A20 blasts [30] (link) in the absence versus presence of TGF-β1. Sixteen hours later, cells were analyzed by flow cytometry and supernatants were analyzed by ELISA.
7
Primary Leukocyte Cell Culture
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Primary leukocytes were cultured in cell growth medium in the presence of indicated cytokines. Cell growth medium contains 45% RPMI 1640, 50% AIM V CTS (Gibco by Life Technologies), and 5% non-AB pooled human serum (Gemini Bio-Products) by volume and was supplemented with 65 ug/ml gentamycin (Quality Biological), and 1X penicillin-streptomycin-glutamine solution (Gibco by Life Technologies).
8
Cell Line Characterization and Culturing
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Non-transformed (208F) and v-src-transformed (208Fsrc3) rat fibroblast cell lines described before7 (link),23 (link) were used in the passage range 18–30. Cells were grown in Eagle’s minimum essential medium (MEM) (Sigma, UK) supplemented with 5% FBS (Sigma, UK), 2 mM l -glutamine and 50 mg/ml penicillin–streptomycin in a penicillin–streptomycin-glutamine solution (Gibco, UK). The cultures were incubated at 37 °C in a 5% CO2/air gassed incubator.
9
Maintenance and Use of SLK and SLKK Cells
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Human KS-derived SLK and SLKK cells (also known as SLK+rKSHV.219) [51 (link),81 (link),94 (link),95 (link)] were a gift from Dr. Don Ganem (UCSF, CA). They were expanded on receipt, frozen in liquid nitrogen, and stored in a cryogenic tank until used in the experiments described hereafter. Cells were thawed and maintained in Dulbecco’s Modified Eagle medium supplemented with 10% v/v fetal bovine serum (Sigma-Aldrich, St Louis, MO) and 1% Penicillin/streptomycin/glutamine solution (Gibco, Carlsbad, CA). Additionally, KSHV-positive SLKK cells were periodically grown under selection with 10 μg/mL puromycin to maintain the viral episome. Not counting the time during which cells were frozen, SLK cells had been kept in culture for less than 3 months before being used for RNA isolation (RNA-Seq), transfection assays, or de novo KSHV infections. SLK cells infected with recombinant rKSHV.219 (SLKK cells) were originally selected over time using puromycin in order to reach a high latent viral expression. Therefore, again not counting the time that they were frozen, the SLKK cells were kept in culture for a longer period of time (6 months to a year) before being used for RNA isolation (RNA-Seq) and transfection assays. All experiments were done in triplicate independent cell cultures maintained at 37°C in humidified 5% CO2.
10
Isolation and Characterization of Primary Human Trabecular Meshwork Cells
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