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Cells in monolayer culture were cultivated at sub-confluence before being washed twice with PBS.
Cells were cultured at sub-confluence and utilized between passages 5 and 10.
Cells were seeded in 6-well plates and grown to sub-confluence.
When the clones reached sub-confluence, they were detached by trypsin/EDTA treatment and expanded.
Cells were lysed at sub-confluence or 2 days post-confluence, as described in the text.
MDA-MB-231 cells were cultured to sub-confluence in 6-well plates.
For experiments, fibroblasts were grown to sub-confluence in medium containing 10% FCS.
After reaching 70 80% sub-confluence, we started the micromass culture of scleral cells.
Cells were grown at 37°C in a humidified atmosphere containing 5% CO2 and maintained at sub-confluence to proliferate.
For cytotoxicity assay, cells were seeded in 24-well chambers and grown for 24 hours to sub-confluence.
Cells were seeded onto 35-mm plates in serum-containing medium until sub-confluence and then transferred to SFM.
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