Penicillin streptomycin

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Penicillin-Streptomycin is a stable, sterile liquid solution containing the antibiotics penicillin and streptomycin. It is commonly used as a supplement in cell culture media to prevent bacterial contamination.

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Penicillin (11 citations) Streptomycin (9 citations)

6 protocols using penicillin streptomycin

Generation and Validation of HCT116 Core-Clock KO Cell Lines

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HCT116 cells (ATCC^® CCL-247™) were cultured in Dulbecco’s Modified Eagle Medium DMEM (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% FBS (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) and 1% Penicillin−Streptomycin (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) in a humidified atmosphere containing 5% CO₂ at 37 °C. For live-cell bioluminescence recording, cells were maintained in phenol red-free DMEM (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) containing 10% FBS, 1% Penicillin−Streptomycin and 250 µM D-Luciferin (Bio-Rad laboratories, Hercules, CA, USA). Cell counting and morphology analysis were performed in a LUNA™ Automated Cell Counter (Logos Biosystems, Anyang, South Korea).
HCT116 core-clock KO cell lines were generated as previously described [42 (link),43 (link)]. In short, HCT116 WT cells were transfected with CRISPR-Cas9 plasmids and guide RNAs targeting core-clock genes BMAL1, PER2 or NR1D1. Positive cells were single-cell sorted based on GFP expression, expanded and validated for KO efficiency on DNA, RNA and protein level.

Fuhr L., Basti A., Brás T.S., Duarte M.F, & Relógio A. (2022). Antiproliferative Effects of Cynara Cardunculus in Colorectal Cancer Cells Are Modulated by the Circadian Clock. International Journal of Molecular Sciences, 23(16), 9130.

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Culturing Human Microglia and Macrophages

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Human microglia clone 3 (HMC3) cells were obtained from the American Tissue Culture Collection (ATCC). Cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM; Gibco, #11995-065), supplemented with 5% fetal bovine serum (FBS; Wisent #080-450) and 1% penicillin–streptomycin (P-S; Invitrogen, #15140-122). THP-1 human monocyte cells were obtained from ATCC. THP-1 cells were cultured in Roswell Park Memorial Institute 1640 (RPMI 1640; Gibco, #11835-030) containing 10% (FBS), 1% penicillin–streptomycin (P/S), and 50 µM of β-mercaptoethanol (Bio-Rad, #161-0710) in a 75 cm² vented cell culture flask (Sarstedt, 83.3911.002). Differentiation of THP-1 monocytes into macrophages was induced over a period of 5 days with phorbal-12-myristate-13-acetate (PMA; Abcam, ab120297) at a concentration of 75 ng/mL. Differentiating THP-1 cells were refreshed with media containing PMA on the third day. HMC3 cells were kept at passages below 35, while THP-1 passages were limited to 12. Both cell types were incubated at 37 °C, 5% CO₂, and atmospheric O₂.

Migliavacca M., Correa-Paz C., Pérez-Mato M., Bielawski P.B., Zhang I., Marie P., Hervella P., Rubio M., Maysinger D., Vivien D., del Pino P., Pelaz B., Polo E, & Campos F. (2024). Thrombolytic therapy based on lyophilized platelet-derived nanocarriers for ischemic stroke. Journal of Nanobiotechnology, 22, 10.

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Dendritic Cell Generation from Bone Marrow

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Bone-marrow-derived dendritic cells (BMDCs) were generated with recombinant human Flt3 ligand (rh FLt3L/CD135) (ImmunoTools, Germany). In brief, bone marrow cells were isolated from the indicated experimental animals as described above and were grown at a concentration of 1×10⁶ cells per well onto a 24-well plate in RPMI 1640 medium supplemented with 10% FCS, 1% penicillin/streptomycin, 50 μM 2-mercaptoethanol (Bio-Rad Laboratories, Germany), and 200 ng/mL rh FLt3 ligand. On day 6, the medium including 200 ng/mL rh FLt3 ligand was renewed. On day 9, the purity of bone-marrow-derived dendritic cells was >90% based on the proportion of the population expressing CD11c as determined by flow cytometry.

Schneble D., El-Gazzar A., Kargarpour Z., Kramer M., Metekol S., Stoshikj S, & Idzko M. (2023). Cell-type-specific role of P2Y2 receptor in HDM-driven model of allergic airway inflammation. Frontiers in Immunology, 14, 1209097.

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Circadian rhythm monitoring in HCT116 cells

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For live-cell bioluminescence recordings, 2.5 × 10⁵ HCT116 cells were seeded in 35 mm dishes and maintained in phenol red-free DMEM (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) containing 10% FBS, 1% Penicillin-Streptomycin supplemented with 250 µM D-Luciferin (Bio-Rad laboratories, Hercules, CA, USA). Cells were synchronized by adding fresh medium prior to measurement and treated with C. cardunculus or a vehicle control where appropriate. BMAL1-promoter-(BLP)-reporter activities were measured, using a LumiCycle instrument (Actimetrics, Wilmette, IL, USA) for five consecutive days. Raw luminescence data were de-trended using the 24 h running average (divided values) using Chronostar analysis software V3.0 (Kramer lab, Berlin, Germany) [45 (link)]. The first 12 h of measurement were removed from the analysis, since the first data collection is comparatively very noisy due to technical limitations of the device. The phase in radian was calculated using the following equation with φ(h) = phase (in h), T = period:

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Circadian Rhythm Monitoring in Colorectal Cancer Cells

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HCT116 cells (ATCC^® CCL-247™) and SW480 cells (RRID:CVCL_0546) were cultured in Dulbecco’s Modified Eagle Medium DMEM (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% FBS (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) and 1% Penicillin−Streptomycin (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) in a humidified atmosphere containing 5% CO₂ at 37 °C. Stable-transduced cells were selected and maintained in medium containing 150 μg/mL hygromycin B (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) for the BMAL1:LUC hygromycin (BLH), 10 μg/mL of blasticidin S HCl (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) for the PER2:LUC blasticidin (PLB) and 1.5 μg/mL of puromycin (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) for the shRNA KD of the clock genes. For live-cell bioluminescence recording, cells were maintained in phenol red-free DMEM (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) containing 10% FBS, 1% Penicillin−Streptomycin and 250µM D-Luciferin (Bio-Rad laboratories, Hercules, CA, USA). Cell counting and morphology analysis were performed in LUNA™ Automated Cell Counter (Logos Biosystems, Anyang, South Korea). Cell lines were tested for mycoplasma by using the Mycoplasmacheck service of Eurofins Genomics (Eurofins Genomics, Ebersberg, Germany).

Basti A., Fior R., Yalҫin M., Póvoa V., Astaburuaga R., Li Y., Naderi J., Godinho Ferreira M, & Relógio A. (2020). The Core-Clock Gene NR1D1 Impacts Cell Motility In Vitro and Invasiveness in a Zebrafish Xenograft Colon Cancer Model. Cancers, 12(4), 853.

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Oxidative Stress Profiling in H9c2 Cardiomyoblasts

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Embryonic ventricular rat heart derived H9c2 cardiomyoblasts (CRL-1446) were from American Type Culture Collection (Manassas, USA); Dulbecco's Modified Eagle's Medium (DMEM), Penicillin-Streptomycin, Phosphate Buffered Saline (DPBS), and fetal bovine serum (FBS) were from Carlo Erba (Milano, Italy); Bradford and RC DC protein assay kit was from Bio-Rad Laboratories (Hercules, USA); 2', 7'-dichlorfluorescein diacetate (DCFH-DA) and MitoSOX fluorescent dyes were from ThermoFisher Scientific (Waltham, USA); ViaCount was from Merck-Millipore (Darmstadt, Germany). Seahorse XF96 microplate plates, Seahorse XF Assay media, Seahorse XF base media without phenol red and Mito Stress Kits were from Agilent (Santa Clare, USA). All other chemicals including palmitate and 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) (MTT) were obtained from Sigma-Aldrich (St. Louis, USA).

Dludla P.V., Silvestri S., Orlando P., Mazibuko-Mbeje S.E., Johnson R., Marcheggiani F., Cirilli I., Muller C.J.F., Louw J., Chellan N., Obonye N., Nkambule B.B, & Tiano L. (2020). Palmitate-induced toxicity is associated with impaired mitochondrial respiration and accelerated oxidative stress in cultured cardiomyocytes: The critical role of coenzyme Q_9/10. Toxicology in vitro : an international journal published in association with BIBRA, 68.

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