Hi Tasfia,

We use RPMI with glutamine and 20% FBS for resuscitation of frozen THP-1 cells, we don't use mercaptoethanol. Then, after the first passage, once the culture is established, we use 10% FBS for successive passaging.

We culture them in T75 culture flasks with vented cap in horizontal position (they should grow in suspension). At the beginning we check them daily until the cells reach the exponential phase of growth. To keep the cells in exponential growth we maintain the cultures between 3-8x10^5 cells/mL.

Another important note with these cells is to avoid DMSO as cryoprotectant since DMSO induces differentiation of THP-1 cells. For cryopreservation we use 90% serum and 10% Glycerol.

THP-1 cells should grow fine in suspension, adherence is usually a sign of suboptimal culture conditions or activation. For passaging, just dilute the cells into new flasks with fresh media, without centrifugation. We passage them when they reach 8-9x10^5 cells/mL.

I hope this helps. Let me know if you have any further questions.

Thank you very much Jose, that certainly helped! I will surely let you know if I have any further questions. Thanks.

Do you know if the centrifugation before passaging can affect the growing of this cells?