How can I get RAW264.7 cells to differentiate into osteoclasts?

Melisa Ait Kaci @Melisa-Ait-Kaci

13 June 2024 1 5K Report

I can't get RAW264.7 cells to differentiate into osteoclasts.

I've tested different densities: 5000 cells/cm2 , 10,000 cells/cm² and 20,000 cells/cm² in 35mm polystyrene.

I use DMEM medium, 10% FBS and 50ng/ml RANKL.

Number of passages: P6-P8

Confluence: 60%.

The medium is changed every 2-3 days by restimulating with 50ng/ml RANKL.

After 5 days, I obtain undifferentiated cells, and on DAPI/Actin labeling, the cells seem to superimpose on each other rather than fuse.

Can anyone help me, please?

Thank you in advance.

Ellen A G Chernoff

Try changing the medium every other day to maintain the RANKL stimulation.

Article Evaluation of culture conditions for osteoclastogenesis in R...

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