I am growing BV2 cells in two different countries from different original stocks but exactly the same medium type and recipe, and the same flasks. In one country the cells do as expected, within 24 hours they're adhering and looking perfectly perky. In the other country 24 hours doesn't seem to be enough for them to adhere, they take up to two or three days sometimes and even then will only adhere to PLO coated flasks, which I've never had to use with this line before. I have absolutely no idea what's going wrong with these cells since I appear to be doing exactly the same in both countries! My only thought is that they're from different stocks BUT I have seen the badly behaved ones grow previously...just not recently!
This is straight from thawing....they're just not coming back to life after i thaw them. I could check viability as soon as i thaw but i'm not sure how sensible that would be?
Dear all,
I am facing a similar problem when thawing BV-2 microglia cell line and I would highly appreciate your help since the only BV-2 cells I have are the ones in culture and I bought them to ICLC. I would rather recover the ones I have now than buying a new vial, if possible.
Two days ago I thawed the only BV-2 microglia vial in RPMI media since it was frozen in that media. 24 hours later I realized that around 30% of the cells were not attached, then I centrifuged the supernatant and I seeded the cells in DMEM, RPMI and DMEM:F12. From the cells I collected from the supernatant, I realized that there are more attached cells in DMEM or DMEM:F12 compared to RPMI media condition. I am not sure if this may be due to the fact that RPMI is also used when cells are grown in suspension (and therefore those cells are viable), or which the cause might be. Is that normal to have cells in suspension in BV-2 cultures? Or should I be worried about? Can it be due to an inflammation?
I kept the attached cells in RPMI media and the vast majority of them are still attached.
My main concerned is the fact that the vast majority of my different plates (attached cells from the very beginning, supernatant maintained in DMEM, DMEM:F12 and RPMI) do not seem to have elongations. I would like to know if there is any way to recover these cells or if it takes some days for the cells to get those elongations. Do you think is worth doing adding primocin or plasmocin to the media so I could get rid of any potential micoplasma contamination that could avoid my cells to get protrusions?
Many thanks for your help!
- Badges
- Science topic