I have read papers and some of them are not suggested to use passage number more than 20. However, my lab has been maintaining the 3T3-L1 and currently at passage 34. Therefore, I curious what is the specific reason which affected the outcome (adipocyte cell) when using passage number more than 20 and should we adjust the amount of reagent used such as insulin, dexamethasone or IBMX to overcome the differentiation efficient even using high passage number. Lastly, do give a conclusion either high passage number of 3T3-L1 might affect the differentiation efficient or not.

Thank you in advance for those who answer this question.