Hello Uma Sriram

Yes, you may harvest BV-2 microglial cells by trypsinization.

BV-2 cells grow as a mixture of floating and adherent cells. You just have to treat BV-2 cells with prewarmed 0.25% trypsin- 0.53mM EDTA for not more than 1 minute. Observe cells under an inverted microscope until cell layer begin to detach. Tap the flask for complete detachment of cells from the surface of the substratum. Add complete medium to inactivate trypsin and aspirate cells by pipetting gently. Add the required amount of cell suspension to a new culture vessel and continue the culture at 37 degree C. You could also centrifuge the trypsinized cells at 1000-1200 rpm for 5 minutes which I usually avoid.

Over exposure of cells to trypsin may harm the cells. Also, avoid scraping the cells as it may cause cell damage and eventually lead to cell death.

Best.

Dear Malcolm,

Appreciate your response. We use 5 ml of prewarmed 0.05% Trypsin-EDTA (in T75 flask or 200ul in an 12 well plate and keep for 5 min at 37 deg, tap the flask or plate well and immediately add 20 ml or 1 ml complete medium respectively, to arrest Trypsin action. Take the cells in a 50 ml tube/ 5 ml tube and spin at 1500 rpm for 5 min. For downstream analysis as flowcytometry we need to spin down the cells. How can you proceed without pelleting and resuspending cells and to replate for a desired cell number?

I am wondering in our expts even this mild treatment is causing cell death!!

If you have any suggestions please let me know.

Thanks

Uma