Hi,
I'm new in the use of human monocyte cell line THP-1, and I'm having some trouble in their culture. Although they tend to grow up relatively well, they seem to disapear when i'm trying to pass them to another flask. I'm wondering if the problem can be that I have to spin them down at higher speed (I'm using 1000rpm for 5 min).
Any suggestion?
Thank you very much!
Aiala
Hello Aiala, Use a fresh aliquots of frozen cells, no more than 6 to 10 passages,
Ensure that your culture is not contaminated, THP-1 cells are very sensitive to contamination and mycoplasma are difficult to detect.
Check you PMA (or differenciation agent) solution.
Be sure that you re using 1000 rpm and not g (gravity).
Thanks Hafid, but it is their second passage, and I haven't used the PMA yet... I'll check the speed, just in case.
Best,
Hi Aiala,
First, during cell passage referring g-force (rcf) is more feasible than rpm. Rcf value changes from one centrifuge to another depending on the radius of the rotor even if you enter same rpm . However, rcf is universal. Second, 200-400 g for 5-10 minutes is convenient for THP1 cell line.
I suggest you to check always cell viability with trypan blue or another method and always take minimum inoculation density into consideration and do not let your cell line to exceed maximum density. It might cause a change in expression profile. Do not let your cell line to wait in PBS or cold media too long. This decreases cell viability.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line