Vero E6 cells acquired from the American Type Culture Collection (ATCC–CRL 1587) were maintained in MEM with stable L-glutamine (PAN-Biotech, Aidenbach, Germany) supplemented with 10% FBS (PAN-Biotech, Aidenbach, Germany), penicillin (100 IU/mL), streptomycin (100 μg/mL) and non-essential amino acids (PAN-Biotech, Aidenbach, Germany). For virus isolation, Vero E6 cells were seeded at 300 000 cells per well in a 6-well cell culture plate and left to adhere overnight. The following day, nasopharyngeal swabs were filtered through a 0.22 μm filter and diluted 1:1 with the addition of MEM with stable L-glutamine supplemented with penicillin (200 IU/mL) and streptomycin (200 μg/mL). Further dilutions to final 1:10, 1:40, and 1:100 were prepared in MEM with stable L-glutamine supplemented with 2% FBS (PAN-Biotech, Aidenbach, Germany), penicillin (100 IU/mL), streptomycin (100 μg/mL) and non-essential amino acids. After 1 h long incubation in a 5% CO2 environment at 37 °C, the inoculum was removed and DMEM with 4.5 g/L glucose and stable L-glutamine (PAN-Biotech, Aidenbach, Germany), supplemented with 2% FBS, penicillin (100 IU/mL) and streptomycin (100 μg/mL), in a volume of 2 mL was added. Cell culture was checked daily for cytopathic effect until day 5 post-infection when the supernatant was collected from the positive cultures.
Penicillin
Manufactured by PAN Biotech
Sourced in Germany, United States, France, Austria, United Kingdom
Penicillin is a class of antibiotics derived from the Penicillium fungus. It functions as an antimicrobial agent, inhibiting the growth and reproduction of certain bacteria. The core function of penicillin is to disrupt the cell wall synthesis in susceptible bacteria, leading to their death or inhibition of growth.
Automatically generated - may contain errors
Lab products found in correlation
Streptomycin, by PAN Biotech (196 mentions)
L glutamine, by PAN Biotech (37 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (36 mentions)
Fetal bovine serum (fbs), by PAN Biotech (33 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (26 mentions)
Fetal calf serum (fcs), by PAN Biotech (21 mentions)
Dulbecco modified eagle medium (dmem), by PAN Biotech (19 mentions)
Rpmi 1640 medium, by PAN Biotech (13 mentions)
Streptomycin, by Thermo Fisher Scientific (12 mentions)
Amphotericin b, by PAN Biotech (11 mentions)
Penicillin, by Thermo Fisher Scientific (11 mentions)
Non essential amino acids (neaa), by PAN Biotech (9 mentions)
Fetal calf serum (fcs), by Merck Group (8 mentions)
Fungizone, by PAN Biotech (6 mentions)
Fetal bovine serum (fbs), by Merck Group (6 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (6 mentions)
Sodium pyruvate, by PAN Biotech (5 mentions)
Penicillin streptomycin, by PAN Biotech (5 mentions)
Glutamine, by PAN Biotech (5 mentions)
Trypsin edta, by PAN Biotech (5 mentions)
209 protocols using penicillin
1
SARS-CoV-2 Virus Isolation in Vero E6 Cells
2
Cell Culture Protocols for Pancreatic Cancer
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Human AsPC-1 and MIA PaCa-2 pancreas carcinoma cells were obtained from Prof. Dr. Ulrich Massing, University Hospital Freiburg, Germany, Clinic for Tumor Biology and their identity was confirmed by Short-tandem repeat profiling. AsPC-1 cells were grown in RPMI 1640 medium (PAN Biotech, Aidenbach, Germany) containing 10% (v/v) fetal calf serum (FCS, Sigma Aldrich, Steinheim, Germany), 100 U/mL penicillin, and 100 µg/mL streptomycin (PAN Biotech). MIA PaCa-2 cells were grown in Dulbecco’s Modified Eagle’s Medium (DMEM) (PAN Biotech, Aidenbach, Germany) containing 10% (v/v) fetal calf serum (FCS, Sigma Aldrich, Steinheim, Germany), 100 U/mL penicillin, and 100 µg/mL streptomycin (PAN Biotech). Capan-2 cells were kindly obtained from Prof. Ingo Schmidt-Wolf, University Hospital Bonn, Germany, and cultivated in RPMI 1640 medium (PAN Biotech) containing 10% (v/v) fetal calf serum (FCS, Sigma Aldrich), 100 U/mL penicillin and 100 µg/mL streptomycin (PAN Biotech). All cell lines were incubated at 37 °C in a humidified atmosphere containing 5% (v/v) CO2.
For subcultivation, cell lines were treated with trypsin/EDTA (5 g/L trypsin; 0.2 g/L EDTA × 4 Na, Sigma Aldrich) for 5 min at 37 °C. Mycoplasma check was routinely performed every month and cell identity was validated using a STR profile analysis.
For subcultivation, cell lines were treated with trypsin/EDTA (5 g/L trypsin; 0.2 g/L EDTA × 4 Na, Sigma Aldrich) for 5 min at 37 °C. Mycoplasma check was routinely performed every month and cell identity was validated using a STR profile analysis.
3
Cell Culture and Viral Infection Protocols
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All cell lines (HEK293T, HeLa, Jurkat, THP-1) were purchased from American type culture collection (ATCC: #CRL-3216, #CCL-2, #TIB-152, #TIB-202). HEK293T and HeLa cells were cultivated in Dulbecco’s Modified Eagle Medium (DMEM, Gibco) supplemented with 10% (v/v) fetal bovine serum (FBS, Gibco), 100 U/ml penicillin (PAN-Biotech), 100 µg/ml streptomycin (PAN-Biotech) and 2 mM L-glutamine (PAN-Biotech) (hereafter called DMEM + 3). Jurkat and THP-1 cells were cultivated in Roswell Park Memorial Institute (RPMI, Gibco) 1640 medium supplemented with 10% (v/v) FBS (Gibco), 100 U/ml penicillin (PAN-Biotech), 100 µg/ml streptomycin (PAN-Biotech) and 2 mM L-glutamine (PAN-Biotech). Cells were tested for mycoplasma contamination by polymerase chain reaction (PCR) test and used if negative. Measles virus (MeV, Schwarz strain) was a generous gift from K.-K. Conzelmann (Max von Pettenkofer Institute, Ludwig-Maximilians-University Munich, Germany). Influenza A virus (strain PR8/34 H1N1) and encephalomyocarditis virus (EMCV, EMC strain) were purchased from ATCC (#VR-95, #VR-129B).
4
Cultivation of Human Cancer Cell Lines
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Human MV3 melanoma cells were cultivated in RPMI 1640 medium (PAN Biotech, Aidenbach, Germany) containing 10% (v/v) fetal calf serum (FCS) (Sigma Aldrich, Steinheim, Germany), 100 U/mL penicillin and 100 μg/mL streptomycin (PAN Biotech). EA.hy926 endothelial cells were cultured in Dulbecco’s modified Eagle’s Medium low glucose (DMEM low glucose) (Sigma Aldrich) with 10% FCS, 100 U/mL penicillin and 10 μg/mL streptomycin. MDA-MB-231 breast cancer cells were maintained in DMEM high glucose medium (PAN Biotech) and supplemented with 10% FCS, 1% l -glutamine (PAN Biotech), 100 U/mL penicillin and 100 μg/mL streptomycin. PC-3 prostate cancer cells were cultivated in RPMI 1640 medium containing 10% (v/v) fetal calf serum (FCS), 100 U/mL penicillin and 100 μg/mL streptomycin, 1% l -glutamine and 1% sodium pyruvate (Thermo Fisher Scientific, Waltham, MA, USA). All cells were incubated at 37 °C in a humidified atmosphere containing 5% (v/v) CO2. For subcultivation, MV3, PC-3, MDA-MB-231 and EA.hy926 cells were detached at a confluency of about 90% with trypsin/EDTA (5 g/L trypsin; 0.2 g/L EDTA × tetra sodium, Sigma Aldrich) for 5 min at 37 °C. Test for absence of mycoplasms were performed routinely every month.
5
Culturing SH-SY5Y and HEK293 cells
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Human SH-SY5Y neuroblastoma cells were cultured in D-MEM/F-12 with 10% fetal calf serum (both Thermo Fisher Scientific), 1× MEM non-essential amino acids, penicillin and streptomycin (all Pan Biotech, Aidenbach, Germany). Human HEK293 kidney cells were cultured in D-MEM (Thermo Fisher Scientific) with 10% Cosmic Calf Serum (Cytiva, Marlborough, MA, USA), penicillin and streptomycin (Pan Biotech). Plasmids transfections were performed using Metafectene (Biontex, Munich, Germany, for HEK293), Metafectene Pro (Biontex, for some SH-SY5Y experiments) or Lipofectamine 2000 (Thermo Fischer Scientific, for some SH-SY5Y experiments), according to the manufacturer’s instructions. For one experiment, the proteasomal inhibitor MG132 (MedChemExpress, Monmouth Junction, NJ, USA) was added to 10 μM for 16 h after transfection, with DMSO used as a vehicle control.
6
Maintenance of Human Cancer Cell Lines
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Human AsPC-1 pancreas carcinoma cells and human MV3 melanoma were maintained in RPMI 1640 medium (PAN Biotech, Aidenbach, Germany) containing 10% (v/v) fetal calf serum (FCS, Sigma Aldrich, Steinheim, Germany), 100 U/mL penicillin and 100 μg/mL streptomycin (PAN Biotech). MDA-MB-231 breast cancer cells were cultivated in DMEM (high glucose) medium (PAN Biotech) and supplemented with 10% (v/v) FCS, 1% (v/v) l -glutamine (PAN Biotech), 100 U/mL penicillin and 100 μg/mL streptomycin. PC-3 prostate cancer cells were cultivated in RPMI 1640 medium containing 10% (v/v) FCS, 100 U/mL penicillin and 100 μg/mL streptomycin, 1% (v/v) l -glutamine and 1% (v/v) sodium pyruvate (Thermo Fisher Scientific, Waltham, MA, USA). All cell lines were incubated at 37 °C in a humidified atmosphere containing 5% (v/v) CO2. For subcultivation, all cell lines were incubated with trypsin/EDTA (5 g/L trypsin; 0.2 g/L EDTA × tetra sodium, Sigma Aldrich) for 5 min at 37 °C. Mycoplasm check was conducted every four weeks.
7
Cell Culture Protocols for HIV Research
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HEK293T (77 (link)) and HeLa TZM-bl indicator cells (78 (link)) were maintained in Dulbecco’s modified Eagle’s medium (Thermo Fisher Scientific) supplemented with 100 U/mL penicillin, 100 μg/mL streptomycin (PAN Biotech), and 10% fetal calf serum (FCS; Sigma-Aldrich). Both cell lines were regularly monitored for mycoplasma contamination using the MycoAlert mycoplasma detection kit (Lonza Rockland). Primary CD4+ T cells were cultured in RPMI 1640 containing l -glutamine supplemented with 100 U/mL penicillin, 100 μg/mL streptomycin (PAN Biotech), 10% heat-inactivated FCS, and 5% human AB serum (Sigma-Aldrich).
8
Isolation and Culturing of T Cells and Target Cell Lines
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Peripheral blood mononuclear cells (PBMCs) were isolated from whole blood obtained from healthy donors upon informed consent and approval by the institutional review board using density centrifugation on Lymphoprep (Axis-Shield, Oslo, Norway). Following extraction, PBMCs were cryopreserved and stored at −80 °C until experimental use. T cells were cultured in RPMI 1640 + GlutaMAX (Gibco, ThermoFisher), 100 IU/mL penicillin + 100 µg/mL streptomycin (Pan-Biotech, Aidenbach, Germany), 2 mM HEPES (PAA, GE healthcare), and 10% (v/v) heat-inactivated fetal calf serum (Pan-Biotech, Aidenbach, Germany).
Target cell lines encompassed 293T cells (human embryonic kidney cells that express the SV40 large T antigen) and SKOV-3 human ovarian cancer cells (ATCC HTB77; American Type Culture Collection, Manassas, VA). Caov-3 ovarian cancer cells were a kind gift from the chair of immunology at the Leibniz-institute for immunotherapy. The cells were maintained in DMEM + GlutaMAX (Gibco, ThermoFisher), 100 IU/mL penicillin + 100 µg/mL streptomycin (Pan-Biotech, Aidenbach, Germany), and 10% (v/v) heat-inactivated fetal calf serum (Sigma-Aldrich, St. Louis, MO, USA).
Target cell lines encompassed 293T cells (human embryonic kidney cells that express the SV40 large T antigen) and SKOV-3 human ovarian cancer cells (ATCC HTB77; American Type Culture Collection, Manassas, VA). Caov-3 ovarian cancer cells were a kind gift from the chair of immunology at the Leibniz-institute for immunotherapy. The cells were maintained in DMEM + GlutaMAX (Gibco, ThermoFisher), 100 IU/mL penicillin + 100 µg/mL streptomycin (Pan-Biotech, Aidenbach, Germany), and 10% (v/v) heat-inactivated fetal calf serum (Sigma-Aldrich, St. Louis, MO, USA).
9
Isolation and Culture of Primary Human T Cells
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Peripheral blood mononuclear cells (PBMCs) were purified from blood from healthy donors upon informed consent and approval by the institutional review board (21-2224-101 Regensburg) using density centrifugation on Lymphoprep (Axis-Shield, Oslo, Norway). T cells were cultured in RPMI 1640 medium supplemented with GlutaMAX (Gibco, ThermoFisher, Waltham, MA, USA), 100 IU/mL penicillin, 100 µg/mL streptomycin (Pan-Biotech, Aidenbach, Germany), 2 mM HEPES (PAA, GE healthcare, Chicago, IL, USA) and 10% (v/v) heat-inactivated fetal calf serum (Pan-Biotech, Aidenbach, Germany). The 293T is a human embryonic kidney cell line that expresses the SV40 large T antigen. BxPC-3 is a human pancreatic cancer cell line (ATCC CRL-1687; American Type Culture Collection, Manassas, VA, USA). The cells were cultured in DMEM supplemented with GlutaMAX (Gibco, ThermoFisher, Waltham, MA, USA), 100 IU/mL penicillin, 100 µg/mL streptomycin (Pan-Biotech, Aidenbach, Germany), 2 mM HEPES (PAA, GE healthcare, Chicago, IL, USA) and 10% (v/v) heat-inactivated fetal calf serum (Pan-Biotech, Aidenbach, Germany).
10
Culturing Caco-2 and HaCaT Cell Lines
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Human colon epithelial cells, Caco-2 (accession No 86010202, European collection of authenticated cell cultures (ECACC), Salisbury, UK), were grown in monolayers in Dulbecco’s Modified Eagle’s Medium (DMEM), supplemented with 10% inactivated calcium-free fetal calf serum (Gibco Laboratories, Grand Island, NY, USA), 100 U/mL penicillin/0.1 mg/mL streptomycin (PanBiotech GmbH, Aidenbach, Germany) and 2 mM L- glutamine (PanBiotech GmbH, Germany), at 37 °C and in an atmosphere containing 5% CO2 (CO2 Incubator; Thermo Scientific, Marietta, OH, USA) [28 (link)]. HaCaT (immortalized human keratinocyte) cells (accession No. 300493, Cell Lines Service GmbH, Eppelheim, Germany) were cultured in DMEM medium that was low in calcium (0.03 mM) and supplemented with 10% calcium-free fetal calf serum, 100 U/mL penicillin/0.1 mg/mL streptomycin and 2 mM L-glutamine (PanBiotech GmbH, Germany).
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