DSG-BC1 and DSG-BC2 cells were plated in 100 × 15 mm Petri dishes (Corning Costar) previously marked with lines in the exterior of the lower part of the Petri dish to guide the scratching. Cells were plated with growth medium at a cell density of 40,000 cells/cm2. The monolayers reached confluence within 24 h. The monolayers were scratched using a sterile pipette to generate a linear wound in eight different lines/plate. Cell motility was evaluated every 3 h; micrographs were taken for the eight scratched lines for each cell type. Using the image processing software Image J (public domain, Wayne Rasband NIH), the cell-free areas were outlined, and the surface was estimated in pixel units. Cell motility was evaluated as the reduction in cell-free area every 3 h and reported as average and standard deviation.
Petri dishes
Manufactured by Corning
Sourced in United States, Italy
Corning Petri dishes are circular, shallow, transparent dishes made of rigid plastic or glass. They are primarily used as a standard laboratory vessel for culturing microorganisms or cells.
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Lab products found in correlation
Streptomycin, by Thermo Fisher Scientific (4 mentions)
Penicillin, by Thermo Fisher Scientific (4 mentions)
Fetal bovine serum (fbs), by Merck Group (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Anti flag m2 magnetic bead, by Merck Group (3 mentions)
Cell lysis buffer, by Merck Group (3 mentions)
Cell lysis buffer, by Beyotime (3 mentions)
Bovine serum albumin (bsa), by Merck Group (3 mentions)
Amphotericin b, by Thermo Fisher Scientific (2 mentions)
Insulin, by Merck Group (2 mentions)
Sodium chloride (nacl), by Merck Group (2 mentions)
Gelatin, by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Roswell park memorial institute (rpmi), by Merck Group (2 mentions)
Penicillin streptomycin, by Merck Group (2 mentions)
Disposable culture flasks, by Corning (2 mentions)
Whatman, by Cytiva (2 mentions)
Streptomycin, by Corning (2 mentions)
Penicillin, by Corning (2 mentions)
74 protocols using petri dishes
1
Scratch Wound Assay for Cell Motility
2
Bovine Mammary Epithelial Cell Isolation
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bMECs isolation was performed on udder alveolar tissue from healthy lactating cows as described (Anaya-López et al., 2006 (link)). Cells from passages 2–8 were used in all of the experiments. The cells were cultured in petri dishes (Corning-Costar) in growth medium (GM) that was composed of a DMEM medium/nutrient mixture F-12 Ham (DMEM/F-12K, Sigma), which was supplemented with 10% fetal calf serum (Equitech Bio), 10 μg/ml insulin (Sigma), 5 μg/ml hydrocortisone (Sigma), 100 U/ml penicillin, 100 μg/ml streptomycin, and 1 μg/ml amphotericin B (Invitrogen). The bMECs were grown in 5% CO2 atmosphere at 37°C.
3
HeLa Cell Infection by Gardnerella vaginalis
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HeLa cells (derived from a woman with cervical adenocarcinoma and HPV 18 infection) were grown on Petri dishes (Corning-Costar) in growth medium composed of DMEM/F-12 (Sigma-Aldrich Cat # D9785) supplemented with 10% fetal bovine serum (Sigma-Aldrich Cat # F2442), 100 U/mL penicillin, 100 μg/mL streptomycin (Gibco Cat # 15140122), and 1 μg/mL amphotericin B (Thermo Fisher Cat # 15290018). HeLa cells were grown under a 5% CO2 atmosphere at 37 °C, and 24 h before infection with G. vaginalis, cells were cultured in serum-free medium without antibiotics. On the next day, HeLa cells were infected for 2 and 4 h, with the following multiplicities of infection (MOI); 1:1 and 20:1 bacterium per cell (15 × 106 CFU/mL) under 5% CO2 and at 37 °C. After infection, culture supernatants were collected and used for indirect ELISAs and dot blots.
4
Neuron Incubation with Conditioned Media
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In a different experimental approach, neurons were incubated in CM derived from MC/9 cells subjected to OGD (Fig. 2D). MC/9 cells were plated at the density 7 Â 10 5 cells/well in 60 mm diameter Petri dishes (Costar Corning Incorporate) and subjected to OGD as previously described. At the end of OGD period alone or at the end of OGD followed by incubation in recovery medium, supernatants were collected, samples were sedimented to remove MC/9 cells and particulates, and 500 μL of CM were transferred to the neurons plated in 24-well plates (Nunc-Thermo Scientific).
5
Evaluating Vessel Infiltration in CAM Model
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To assess the influence of the 3DP PCL framework within the CHyA-B matrix on vessel infiltration ex vivo, scaffolds were incubated within a chick chorioallantoic membrane (CAM) model as previously described (do Amaral et al., 2019a (link); Ryan et al., 2019 (link); do Amaral et al., 2019b (link)). Fertilised chicken eggs (Ovagen Group Ltd., Co. Mayo, Ireland) were purchased at day 0 of development and incubated at 37°C for 3 days. Following incubation, the eggs were cracked into 100 mm diameter petri dishes (Corning Inc., New York, United States), which in turn were then placed within larger 150 mm diameter petri dishes containing sterile PBS, to form a humidified chamber. The chicks were incubated for a further 4 days, completing 7 days of total development, viability of embryos was firstly ensured and checked at every day from this point forward; when cross-linked and sterilised CHyA-B matrices and reinforced composite CHyA-B scaffolds were placed within the chick membrane and incubated for a further 5 days, allowing for vessel ingrowth. At day 12 the samples were harvested and imaged. The vascularisation around the scaffolds was quantified using ImageJ software after treatment with the “Mexican Hat Filter” to outline the blood vessels, and then converted to 8-bit and the blood vessel area was measured.
6
C. elegans Maintenance on NGM Plates
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N2 ancestral Bristol strain of
C. elegans (CGC, Minneapolis, MN, USA) were maintained on NGM lite plates (N1005, US Biologicals, Salem, MA, USA) with bacterial lawns at 25±0.5°C for all experiments. Bacterial lawns were grown on Petri dishes (10, 60, 100 mm Corning, USA) overnight using 10, 20 or 60 µl of 1X
E. coli OP50 (CGC, Minneapolis, MN, USA; 1X=OD
600=8.0×10
8 cells/mL).
C. elegans (CGC, Minneapolis, MN, USA) were maintained on NGM lite plates (N1005, US Biologicals, Salem, MA, USA) with bacterial lawns at 25±0.5°C for all experiments. Bacterial lawns were grown on Petri dishes (10, 60, 100 mm Corning, USA) overnight using 10, 20 or 60 µl of 1X
E. coli OP50 (CGC, Minneapolis, MN, USA; 1X=OD
600=8.0×10
8 cells/mL).
7
Isolation and Characterization of Bacterial Membrane Vesicles
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L. reuteri biofilm was developed into 90 mm diameter Petri dishes (Corning Incorporated, New York, NY, USA) as previously described and MVs were isolated from the planktonic and biofilm phenotypes as previously described [15 (link)]. The isolated pMVs and bMVs were quantified by using PFC according to Puca et al. [38 (link)] and used for the evaluation of the antimicrobial activity against the microorganisms aforementioned, as well as for the analysis of the metabolic profiles.
8
Differentiation of iPSCs into Neural Cells
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Patient USC-reprogrammed iPSCs were digested into small clumps using 0.5 mM EDTA. Small clumps were then transferred to Corning Petri dishes and suspended as embryoid bodies (EBs) in iPSC medium for 8 days without basic fibroblast growth factor (bFGF) as previously described (Macarthur et al., 2012b (link)). EBs were then seeded onto cell culture plates in neural induction medium containing DMEM/F12 with Glutamax, 1× NEAA, 1× N2, and bFGF (20 ng/mL). After 2– 3 days, neural rosettes were manually isolated and dissociated into single cells. The cells were expanded in Neurobasal medium supplemented with 1× NEAA, L-Glutamine (2 mM), 1× B27, and bFGF (20 ng/mL).
9
Differentiation of iPSCs into Neural Cells
10
Fibroblast Protein Extraction and Western Blot
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The cells of each fibroblast population were seeded at a density of 1,000 cells/cm2 into 10-cm diameter Petri dishes (Corning, Inc., Corning, NY, USA) and cultured for 7 days (95–100% confluence). The culture medium was refreshed every two days. The cell lysates were harvested according to the standard protocol in NP-40 cell lysis buffer (Thermo Fisher Scientific, Inc.). The supernatant was collected into a fresh microtube containing Protease Inhibitor Cocktail (Sigma-Aldrich; Merck KGaA). The total protein concentration was detected using the Bradford method for protein quantitation (28 ). Samples were resolved by 1-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels according to standard techniques. Equal quantities of total protein (10 µg) were subjected to 12% SDS-PAGE (electrophoresis for 60 min, 100 V, in cold room at 4°C) and transferred onto polyvinylidene difluoride membranes. The membranes were subsequently blocked with 5% goat serum (Sigma-Aldrich) for 1 h and probed with specific SMA primary antibody (dilution, 1:1,000; overnight at 4°C), followed by the appropriate horseradish peroxidase-conjugated secondary antibody (dilution, 1:5,000; 60 min at room temperature). Proteins were detected using the KPL TrueBlue blotting detection reagents (BioVendor Laboratory Medicine, Inc., Brno, Czech Republic).
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