I recently thawed a vial of THP-1 cells . The revival was low but I could see good number of cells growing. But two days later the cell population increased but a lot of cells settled down and now they are differentiated even without a stimulant. I don't see any turbidity in the media. No contamination. Could it be low media volume? Also I left the flask horizontal and not vertical.
Hi Ira Sharma
It's crucial to consider how you culture your THP-1 cells. Higher passages can lead to cell differentiation, which can negatively impact their sensitivity and activity. While some studies suggest that passages up to 25 are acceptable, it's always better to use lower passages (
Dear Dr. Ira Sharma
THP-1 cells are sensitive to seeding density. A high cell seeding density could improve THP-1 characteristics by increasing CD14 expression. CD14 is a leukocyte differentiation antigen that exists on the surface of monocytes/macrophages lineage, especially in macrophages, and is a key marker of pro-inflammatory macrophage, and is often used as an indicator of THP-1 cell differentiation. Its expression can be further increased with the induction of PMA and other inducers.
You may want to refer to the articles attached below for more information.
Article Effect of Culture Conditions on the Phenotype of THP-1 Monoc...
Article Optimization of differentiation and transcriptomic profile o...
Article Optimizing THP-1 Macrophage Culture for an Immune-Responsive...
Article CD14, a Receptor for Complexes of Lipopolysaccharide (LPS) a...
Best.
Hi Ira, immune cells are tricky. Any environmental change can lead to their activation. You may also want to check the culture media ingredients because high glucose can promote cell differentiation. There are many papers available.
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