The cell microenvironment, which is critical for stem cell maintenance, contains both cellular and non-cellular components, including secreted growth factors and the extracellular matrix1,2,3.
Forty-eight hours following cell seeding, the plating medium was exchanged with 30 μL of maintenance medium containing 1 500 penicillin/streptomycin. Maintenance medium was subsequently changed every other day for 12 days.
Finally, half of the tubes are refed with 1 ml of maintenance medium containing normal OS and the other half with maintenance medium containing 5percentt (v/v) rinderpest immune serum.
At day 7, beating cardiomyocytes were observed and the media was changed to cardiac maintenance media containing CDBM and insulin.
After 1-h adsorption of inoculum at 37 °C, inoculum was removed and overlaid with maintenance media containing 0.9% agarose (BD BaculoGold Plaque Assay Agarose).
Throughout that process, aliquots of maintenance medium containing single fibers were transferred onto clean 18 mm-diameter glass coverslips coated with 15 μg cm−2 Matrigel (BD biosciences) in 12-well tissue culture plates.
To test for cytotoxicity, 0.1 mL of the maintenance medium containing the tested samples in serial twofold dilutions was added to each of the wells.
A few years later, Ramsden and Northcote ([1987]) found that TEs could be induced when Pinus sylvestris L. suspension cultures were transferred from PRL-4 maintenance medium containing 2% sucrose, 2 mg L−1 NAA and 2 mg L−1 2,4-D thethe same basal medium supplemented with 6% sucrose, 10 mg L−1 NAA and 2 mg L−1 kinetin.
After 48 h, the medium was replaced with maintenance medium containing DMEM supplemented with 10% FBS.
For hippocampal slices, drug incubation was performed at 30 32°C in submersion maintenance chambers containing ACSF saturated with bubbling 95% O2/5% CO2.
