If you plan to test extracts of samples using indirect system model it is important to treat samples for 24, 48, or any other time you expect to test. If you plan to test samples directly using direct contact system no pre-treatment with extract samples is required in this case because contact direct effect and indirect contact effects should be assessed separately.
May be your question is about which time of extraction is better to be used! If that, the most extraction used time in the literature for cytotoxicity testing is 24h.
It is better to have 24 hour studies. But in some case if u don't find results it means your drug has no interaction or may be no effect then preferred to 48 hr studies.