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Penicillin

Manufactured by Vitrocell
Sourced in Brazil

The Penicillin is a laboratory equipment designed to facilitate the cultivation and handling of penicillin-producing microorganisms. It provides a controlled environment for the growth and maintenance of these cultures.

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5 protocols using penicillin

1

Cultivation of Human Umbilical Vein Endothelial Cells

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Human umbilical vein endothelial cells (HUVEC, American Type Culture Collection [ATCC® CRL-1730]) were cultivated in Dulbecco’s Modified Eagle Medium (DMEM, Vitrocell) supplemented with 10% fetal bovine serum (FBS, Nutricell), penicillin (10.000 U.I./mL) and streptomycin (10 mg/mL) (Vitrocell). Cells were maintained incubated at 37 °C, on atmosphere with 5% CO2. Subcultures of cells were performed as instructed by the supplier, using trypsin-EDTA. Cells were used between passages 8 to 15 and counted on a TC20 automated cell counter (Bio-Rad) using trypan blue stain solution at 0.4% (Thermo Scientific).
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2

Culturing Triple-Negative Breast and Endothelial Cells

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Triple-negative breast adenocarcinoma cells (MDA-MB-231, ATCC® HTB-26™) and human umbilical vein endothelial cells (HUVEC, ATCC® CRL-1730™) were cultured in Dulbecco's modified Eagle's medium (DMEM, Vitrocell) supplemented with 10% (v/v) of fetal bovine serum (FBS, Vitrocell), penicillin (100 IU/ml) and streptomycin (100 mg/ml, Vitrocell), and l-glutamine (2 mM), in a humidified incubator at 37 °C and 5% CO2. HUVECs were maintained from passages 8 to 20.
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3

HaCaT Cell Culture for Lidocaine Assay

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HaCaT cells were cultured similarly to a previous method employed by our group [9 (link),10 (link)]. Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 100 U/mL penicillin, 10% fetal bovine serum, and 100 μg/mL streptomycin (Vitrocell, Campinas, Brazil) were used. The cells were kept under a humidified atmosphere at 5% CO2/95% air at 37 °C. The medium was replaced every 2 days, and the culture was duplicated when cells achieved 70 to 80% confluence. The cells were then rinsed with phosphate-buffered saline (PBS) and treated with trypsin (0.25%) for 5 min. The trypsinization process was neutralized in DMEM supplemented with fetal bovine serum. Cells were centrifuged at 3000 rpm for 5 min (20 °C). The supernatant was removed. Fresh medium was added for the viability assay, and LDC, LDC-Nano, and LDC-Epi (prepared immediately before its use by adding 1:100,000 epinephrine—Sigma-Aldrich, St. Louis, MO, USA) to a lidocaine solution were tested.
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4

In Vitro Screening of Antischistosomal Compounds

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Seven weeks after infection, S. mansoni were removed from the hepatic portal system and cultured in RPMI 1640 culture medium (supplemented with 5% inactivated fetal calf serum and 100 U/mL penicillin and 100 μg/mL streptomycin (Vitrocell, Campinas, SP, Brazil) at 37°C in an atmosphere of 5% CO2 until usage. For the determination of activity against adult schistosomes, all samples were initially tested at the concentration of 100 μM (compounds) or 100 μg/mL (extracts), using DMSO stock solutions (10 mM) diluted in supplemented RPMI 1640 medium within 24 flat bottom well plates (Tissue Culture Plastics, TPP, St. Louis, MO) with a final volume of 2 mL per well [33 (link)]. Samples were tested in triplicate with two worms of both sexes placed into each well. Wells with the highest concentration of DMSO in medium (0.5%) served as controls. Praziquantel (2 μM) served as positive control. Parasites were kept for 48 h (37°C, 5% CO2), and their viability was assessed via microscopic readout (Leica Microsystems, Wetzlar, Germany) [34 (link)]. Next, samples presenting antischistosomal activity were tested at lower concentrations, as described above, and each experiment was performed at least three times [35 (link)].
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5

Colonic Tissue Explant Culture and Cytokine Analysis

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Colonic tissue fragments obtained at the end of the experimental model were washed with sterile 0.9% saline solution so debris were removed and placed in 24-well plates in 1 ml of Dulbecco’s Modified Eagle’s Medium (DMEM) (Vitrocell Embriolife, Campinas, SP, Brazil) containing 10% of fetal bovine serum (FBS) (Vitrocell Embriolife, Campinas, SP, Brazil) and 0.1% of antibiotics (streptomycin, amphotericin and penicillin) (Vitrocell Embriolife, Campinas, SP, Brazil). Explants remained in culture at 37°C under controlled 5% CO2 atmosphere for a 24 h period. Next, the supernatant culture medium was collected and used for quantification of TNFα, IL-10 and metalloproteinase-9 (MMP-9) via enzyme-linked immunoassay (ELISA). Analysis was carried out using BD Opteia® and DuoSet® commercial kits according to manufacturer instructions (BD Biosciences, Franklin Lakes, NJ, United States) (R&D Systems, Minneapolis, MN, United States).
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