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However, these authors used a much higher DMSO concentration and maintained the cells at 10.5 °C, which is obviously different to our experimental setup (37 °C).
We examined the ability of Pyk2 deficient osteoblasts to regulate COX-2 protein levels by exposing osteoblasts to 1 hour of FF followed by a 3 hour post-flow incubation or maintained the cells in static culture conditions.
Microscopy was performed on Zeiss Axiovert 200 using a 20× objective equipped with an automated stage and an environmental control chamber, which maintained the cells at 37°C and 5% CO2.
Moreover, it was not determined whether the surfaces had maintained the cells phenotype.
We maintained the cells in Dulbecco's modified Eagle's medium (Invitrogen, Carlsbad, CA, USA).
Next, we maintained the cells in phenol red-free DMEM with 0.2% charcoal-stripped FBS for 24 h.
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AdMSCs within the 3-D aggregates maintained the cell surface epitopes of AdMSCs with high viability.
We then maintained the cell culture at 37 °C, a temperature best supporting the growth of epithelial cells.
NK maintained the cell cultures and provided technical help.
KC maintained the cell line and helped measuring IL-1α with ELISA.
Considering the need of maintaining the cells inside the microcapsules, fermentation at 37 °C and at neutral pH was favorable.
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