MRC5 human lung fibroblast cells (RRID:CVCL_0440) were purchased from RD Biotech (Besançon, France). Colo320 colon cancer cells (RRID:CVCL_1989) were purchased from ATCC (American Type Culture Collection, Manassas, VA, USA). MRC5 and Colo320 cell lines were cultured in Roswell Park Memorial Institute medium (RPMI 1640) supplemented with 10% of heat inactivated fetal calf serum and 1% penicillin plus streptomycin (Gibco, Illkirch, France). Panc‐1 pancreatic cancer cells (RRID:CVCL_0480) were kindly provided by IGBMC (Institute of Genetics and of Molecular and Cellular Biology, Illkirch, France). BPC8 cell lines were generated in our laboratory from ascites of patients with pancreatic ductal adenocarcinoma. Panc‐1 and BPC8 were cultured in Dulbecco's Modified Eagle Medium (DMEM; Gibco) supplemented with 10% of heat‐inactivated fetal calf serum and 1% penicillin plus streptomycin (Gibco). Cells were cultured at 37 °C in a humidified atmosphere of 5% CO2 in air. All cells were periodically authenticated by morphologic inspection. All cell lines were routinely tested for Mycoplasma by PCR.
Penicillin plus streptomycin
Manufactured by Thermo Fisher Scientific
Sourced in United States
Penicillin plus streptomycin is a commonly used antibiotic mixture for cell culture applications. It provides broad-spectrum antimicrobial activity to prevent bacterial contamination in cell culture media and experiments.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (7 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (6 mentions)
Rpmi 1640, by Thermo Fisher Scientific (4 mentions)
Thp 1, by American Type Culture Collection (2 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (2 mentions)
Opti mem, by Thermo Fisher Scientific (2 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (2 mentions)
Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions)
Ml385, by Selleck Chemicals (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 3000 transfection reagent, by Thermo Fisher Scientific (1 mentions)
C57bl 6 background, by Charles River Laboratories (1 mentions)
Mgc 803, by National Collection of Authenticated Cell Cultures (1 mentions)
Bgc823, by National Collection of Authenticated Cell Cultures (1 mentions)
Sirna wizard software 3, by InvivoGen (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Hk 2 cells, by Thermo Fisher Scientific (1 mentions)
Lipopolysaccharide lps, by Merck Group (1 mentions)
Nci h292, by American Type Culture Collection (1 mentions)
E myco mycoplasma pcr detection kit, by iNtRON Biotechnology (1 mentions)
17 protocols using penicillin plus streptomycin
1
Cell Culture Protocols for Cancer Research
2
Culturing Rat Cardiomyocyte H9c2 Cells
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The rat myocardial cell line, H9c2 cells, was used in this study. Dulbecco's modified eagle medium (Gibco, Rockville, MD, USA) containing 10% fetal bovine serum (Gibco, Rockville, MD, USA) and 1% penicillin plus streptomycin (Gibco, Rockville, MD, USA) was used to culture H9c2 cells. H9c2 cells were cultured in an incubator with 37°C and 5% CO2.
3
Establishing H9c2 Hypoxia-Reoxygenation Model
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The rat cardiomyocyte cell line H9c2 was purchased from Shanghai Saibaikang Biotechnology Co. (Shanghai, China). We cultured cells in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, Rockville, MD, USA) containing 10% fetal bovine serum (FBS) (Gibco, Rockville, MD, USA) and 1% penicillin plus streptomycin (Gibco, Rockville, MD, USA). The cells were cultured in an incubator at 37°C and 5% CO2. ML385 (Selleck, Shanghai, China), a nuclear transcription-related factor 2/heme oxygenase-1 (Nrf2/HO-1) signaling pathway inhibitor, was used to inhibit the Nrf2/HO-1 signaling pathway in H9c2 cells. The H9c2 cell hypoxia-reoxygenation model was constructed as follows: After the H9c2 cell density reached 90%, we replaced the cell medium with phosphate-buffered saline (PBS). After filling the incubator with 95% N2, we placed the cells in an incubator for 4 h under hypoxia. Then, we removed the PBS buffer and replaced it with DMEM complete medium [12 (link)].
4
Cardiac Cell Line Injury Model
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The rat myocardial cell line (H9c2) used in this study was obtained from the National Institutes of Health. We used Dulbecco's Modified Eagle's Medium (Gibco, Rockville, MD, USA) containing 10% fetal bovine serum (Gibco, Rockville, MD, USA) and 1% penicillin plus streptomycin (Gibco, Rockville, MD, USA) to culture H9c2 cells. H2O2 was used to induce H9c2 cell injury.
5
Overexpression of ApoE in Cell and Mouse Models
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MFC, MC38, THP-1, MGC-803 (MGC), and BGC-823 (BGC) were obtained from the Cell Bank of the Chinese Academy of Sciences, and tests free for mycoplasma. Cells were maintained in DMEM/F12 with 10% fetal bovine serum (Gibco) and penicillin plus streptomycin (Gibco). The plasmid of ApoE overexpression (ApoE-OE, PGMLV-4931-Apoe) and respective control vectors (PGMLV-4931) were provided by Shanghai Genomeditech Company and were transfected using Lipofectamine 3000 Transfection Reagent according to the manual (ThermoFisher). Female 8-week-old wild-type and Apoe−/− mice with a C57BL/6 background were purchased from Vital River Laboratory Animal Technology Company (Beijing, China). All animal experiments were carried out according to the Principles of Laboratory Animal Care and approved by the Ethics Committee of Xinhua Hospital, Shanghai Jiao Tong University.
6
Silencing LINC00664 in Glioma Cell Lines
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The human glioma cell lines U87 and U251 were purchased from ATCC (Manassas, VA). Cells were maintained in DMEM (Gibco, USA) supplemented with 10% fetal bovine serum (Gibco, USA) and 1% penicillin plus streptomycin (Gibco, USA) and incubated in a humidified incubator (37 °C, 5% CO2).
The sequences of siRNA were designed based on the NCBI Reference Sequence: NR_037194.1 using the siRNA Wizard software 3.1 (InvivoGen). Sequences of siRNAs were as follows. LINC00664 siRNA #1 GGTGATGACAGAATTGTAACA. siRNA #2 GTGACTGTTCTATTCATCATA. siRNA #3 GTTCAAATGGGACATATCTTA. Si-NC GAGGCGAATCGAATTAGATAT. LINC00664 siRNAs were transfected by using Lipofectamine 2000 (Invitrogen, USA) according to the manufacturer’s instructions.
The sequences of siRNA were designed based on the NCBI Reference Sequence: NR_037194.1 using the siRNA Wizard software 3.1 (InvivoGen). Sequences of siRNAs were as follows. LINC00664 siRNA #1 GGTGATGACAGAATTGTAACA. siRNA #2 GTGACTGTTCTATTCATCATA. siRNA #3 GTTCAAATGGGACATATCTTA. Si-NC GAGGCGAATCGAATTAGATAT. LINC00664 siRNAs were transfected by using Lipofectamine 2000 (Invitrogen, USA) according to the manufacturer’s instructions.
7
MLE-12 Cell Culture and LPS Injury
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The mouse alveolar epithelial cell line (MLE-12 cell) was used in the present study. Dulbecco’s modified eagle medium (DMEM) (Gibco, Rockville, MD, USA) containing 10% fetal bovine serum (Gibco, Rockville, MD, USA) and 1% penicillin plus streptomycin (Gibco, Rockville, MD, USA) used to culture MLE-12 cells in an incubator at 37°C and 5% CO2. LPS was used to induce MLE-12 cell injury.
8
Culturing Glioma and Microglial Cells
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Mouse microglioma cell (GL261), mouse microglial cells (BV-2) and human glioma cells (U251) were purchased from the Cell Bank of the Chinese Academy of Sciences. These cells were grown in DMEM (Gibco, USA) medium with a high glucose content, 10% fetal bovine serum (Gibco, USA), and 1% penicillin plus streptomycin (Gibco, USA), all in a humidified environment of 5% CO2 at 37 °C.
9
Stimulation and Damage of HK-2 Renal Cells
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HK-2 cells were purchased from Shanghai Yaji Biotechnology Co. (Shanghai, China). Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, Rockville, MD, USA) was used to culture HK-2 cells. We prepared 10% fetal bovine serum (FBS) (Gibco, Rockville, MD, USA) and 1% penicillin plus streptomycin (Gibco, Rockville, MD, USA) in DMEM to make complete medium. The cells were cultured in an incubator at 37°C and 5% CO2. When the cell growth density was about 60%, we added or transfected the cells. Lipopolysaccharide (LPS, Sigma, St. Louis, MO, USA) was used to induce HK-2 cell damage, while CGRP (Invitrogen, Carlsbad, CA, USA) was used to stimulate HK-2 cells to detect their effects.
10
Cell culture and stimulation protocol
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Human airway epithelial cell line NCI-H292 and human monocytic leukemia cell line THP-1 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). Cells were grown in RPMI 1640 supplemented with 10% fetal bovine serum (Gibco Inc., Brooklyn, NY, USA) in the presence of penicillin plus streptomycin (Gibco Inc.), and 25mM of HEPES at 37 °C in a humidified with 5% CO2. Cells were split every three or four days and were used within 20 passages. All cell cultures were tested and determined to be free of mycoplasma contamination (e-Myco™ Mycoplasma PCR Detection kit, iNtRON Biotechnology, Seongnam, Korea). Cells at a confluence of 60-80% were stimulated with the experimental reagents in serum-free medium as indicated in each experiment.
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