I am treating human PBMCs with IL-4 and anti-CD40 antibody to induce class-switching of B cells in vitro, a protocol widely used in the literature. The incubation period range from 5-7 days (if detecting IgE using ELISpot) to 12-14 days (to detect IgE using ELISA).
It is unclear to me whether media should be changed/replenished, or whether the cells should be split during these extensive incubations (methods from the literature do not mention anything), can anyone with experience clarify whether I should leave the cells alone for the duration of the incubation, or if it will be necessary to change/replenish the media over the course of the incubation?
- Media Change: Perform media changes every 2-3 days during extended incubations to maintain cell health and experimental conditions.
- Cell Splitting: Usually not necessary unless high cell density is observed, which is uncommon in IgE class-switching protocols.
These practices will help ensure the cells remain in optimal conditions for IgE production and provide more reliable experimental results.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line