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On the second day, the culture media were replaced by 2% FBS-DMEM (minimum growth media).
The culture media were replaced and drugs were added every 5 days.
After 16 h, media were replaced with 5 ml of fresh media containing 107 BMNCs/flask.
Media were replaced with neurobasal growth media following aliquot removal.
Media were replaced every 4 days and media changes were performed in normoxic conditions.
Media were replaced 24 h later with complete media containing the indicated concentrations of tamoxifen (Sigma).
Media were replaced twice per week with freshly-prepared regular or osteogenic media.
At a regular time interval, the liquid media were replaced by aliquots of fresh media.
Whole media were replaced with fresh media at specific time intervals.
Further, the media were replaced by 3- 4,5-dimethylthiozolyl-2 -2,5-diphenyl tetrazolium bromide (MTT) (5 mg/mL).
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The media were replaced with fresh DMEM and the cells were cultured for 3 days.
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