Being a suspension cell line why do I see some of the cells getting attached to the flasks? Since this is the first time that I am working with these cell lines, can anyone please help me? I am using RPMI-1640 medium for cell culturing.
Thanks.
Hi,
This previous post can help you:
https://www.researchgate.net/post/Culturing_THP-1_cells
Good luck
THP-1 cell line or any suspension cell line when gets attached to bottom of flask might be due to the death of the cells. If you are observing this attachment during the proliferation then there is a high chance that the cells are not viable. You can determine this simply by using trypan blue exclusion method.
You can maintain this cultures between 2-9x105 cells/ml for avoiding the adherence
There are several reasons for of a small population suspension cells to adhere to the flack bottom. 1. The cells population is heterogeneous and thus a small percentage may express more adhesion molecules. 2. As a result of production, the surface of the flasks may small areas with slightly different charges that may allow cells to adhere. 3. THP-1 cells were derived from a patient with acute monocytic leukemia (monocytes). It’s not unusual for a small percentage of THP-1 cells to spontaneously differentiate into macrophages that will adhere to the flask.
Spontaneous differenciation in macrophages is the most probable. But if you want to keep them in suspension, you can let stand them inn flasks in the vertical position. Just a few cells will adhere onn the cell-culture treated surface.
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