Q1: Why are almost no lipid droplets visible after 2 weeks of induction?
This may be due to a high cell passage number, poor cell health, or insufficient seeding density. Ensure you use cells with a passage number of less than 8, and initiate induction only when the cell confluence reaches nearly 100%.
Q2: What should I do if only a few small lipid droplets appear, and they fail to fuse and enlarge?
Improper storage of the culture medium can lead to the degradation of its components, preventing lipid droplets from fusing and growing. Ensure that the induction period is maintained for the full 14 to 21 days.
Q3: How can I resolve the issue of cells proliferating but failing to differentiate?
Cells that proliferate without differentiating are likely the result of an insufficient cell density prior to induction; adipogenic differentiation requires a high cell density to successfully initiate the process.
Q4: Why do cells experience massive die-off 1 to 3 days after adding the induction medium?
Cell death can occur if the culture medium is not pre-warmed, if the medium has been stored improperly, or if the induction and maintenance media are applied in the incorrect sequence. Always pre-warm the culture medium to 37°C before beginning any procedures.
Q5: Why is the Oil Red O staining very faint, showing almost no coloration?
Faint staining is typically caused by an insufficient number of lipid droplets or a staining duration that is too short. Consider appropriately extending both the induction period and the staining time.
Q6: Can the culture medium still be used if a white precipitate appears?
A white precipitate may indicate the precipitation of additives; if it dissolves completely after warming to 37°C, the medium can be used as normal. However, if the medium appears heavily turbid, discolored, or contains substantial precipitates, it is likely contaminated or has expired and should be discarded immediately.