I read in some articles that after isolation of primary monocytes I must culture them for a couple of hours or overnight and after that I can stimulate them with LPS. Why? For adherence? The monocytes will be stimulated more? Or can I stimulate them immediately after isolation?
The optimal culture time for primary monocytes after isolation with CD14+ microbeads can depend on your specific experimental goals and the downstream applications. In general, allowing the isolated monocytes to recover for 24-48 hours in a suitable culture medium before stimulation with lipopolysaccharide (LPS) is common. This period helps them to regain viability and functionality.
However, the exact duration can vary based on factors such as the isolation method, cell purity, and the specific requirements of your experiment. It's advisable to refer to literature related to your experimental setup or consult with colleagues experienced in similar protocols.
Muhammad Umar thank you for your answer! I would like to stimulate them with LPS during 4,6 and 24h or Pam3CSK. It's for RT qPCR analyzing or ELISA in the surnageant. In the literature, generally, they isolate monocytes to transform them into macrophages. Some articles say that you can culture them during 3h or overnight for stimulating them wtih LPS but I have read these in 2 articles only.
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