For the last two times, I have had near 1 million cells after isolation, they attach within a day, they start to proliferate, but after 3-5 days stops dividing, enter apoptosis and detach.
Do you have any idea what could be the problem?
I would check for contaminations and the media pH. Are your mesenchymal derived from bone marrow? Because sometimes it is source related, such as donnor age....
Thank you all. They are adipose tissue derived stem cells. I used collagenase type 1.
I would check the mycoplasma contamination, i encountered the same trouble previously. The M-plasmocin from invitrogen can work well in my culture system. the drug could be added into medium directly once the cells are seeded.
I too face similar problem. Am isolating MSC's from rat bone marrow. Am getting well attached proliferating cells. I trypsinised it into next passage. Cells were growing fastly and was almost 80% confluent on second day. But the next day, the cells were seen floating with some cells still attached( but they seems to be granular).
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