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Confluent Vero cells in a 96-well tissue plate were inoculated in triplicate with virus suspension (50 μL) [HSV/Blue, at multiplicity of infection (MOI) 1] and culture medium (50 μL) containing testing compounds at different concentrations.
Cells were grown overnight on 12 mm circular glass slides in conditioned medium in a sterile 24-well tissue plate until they reached ~70% confluence as previously optimized in the live cell sialidase assay [29],[29].
In addition, the Peptide-G treated surface, due to the presence of RGD on the surface of the graphene, significantly accelerated the proliferation of human mesenchymal stem cells (hMSCs) at a better rate regarding the tissue plate.
At this early stage of contact inhibition, we noticed that a small number of spherical-shaped cells appeared to be tethered to areas of the tissue plate that were not covered with flattened contact inhibited cells.
The HepG2 cells were seeded in 6-well tissue plate and grown to 80 90% confluence prior to infection.
Cartilage pieces weighing a total of approximately 200 mg were placed into each well of a 24-well tissue plate with 1 ml/well of DMEM supplemented with 10% FBS.
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hMSC were seeded in 12-well tissue plates 1 or 2 days before setting up the co-cultures.
Detached cells were collected and plated on either 24-well tissue culture plate slides or in 6-well tissue plates at a density of 67,500 cells per cm2 and maintained at 37°C for 1 hour.
Cells were seeded at a concentration of ∼2×105 cells per well in 12-well cell tissue plates and monolayers were used 24 h after seeding.
For acute LPS stimulus, cells were harvested from tissue plates and stimulated as previously described.
For transient transfection studies, cells were seeded into 24-well tissues culture plates for HEK293T or 12-well tissue plates for tammar primary cells at optimal densities.
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