I am wondering how many days after MSC isolation from bone marrow you wait until they attach the surface. I isolated them 1.5 days ago and nothing attached yet!! I have a lot of cell an it seems a little weird because I am not supposed to collect lots of them. They might be blood cells but still I think I should have MSCs among them as well.
Hi Sahar
You should be able to see a few adherent fibroblastic-like cells within a few days. If you have a lot of non-adherent cells it would be best to remove half the medium and gently replace with fresh medium.
Regards Rosemary
Working with human bone marrow, we don't even look at the flasks for 4 days post plating to allow cells to adhere. Even after this point it can be another few days before you can identify colonies
While working with BM -MNCs after 48-72 hours , remove the non adherent cells floating in the media , few fibroblast like plastic adherent cells will appear , then change media twice weekly , confluent colonies may appear from next week.
Agree with all these comments. Just few questions: are you working with human or animal BM? Are you remove red cells in some way like osmotic shock or gradient centrifugation? Make sure working with gas-treated culture flasks for adherent cells.
Lots of blood cells could mask the adhered cells, and working with human cells you should wait from 4 to 5 days before adhered cells elongate in a MSC-like morphology.
Good luck
Thank you for the answers. I am working with human cells and used centrifuge to remove blood cells.
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