I am having problem growing my 3T3-J2 which had a change of morphology after few passages? The cells were fine a day after thawing and looked healthy but started to have morphology changes and not growing in number even after a week of culturing.
A sudden change in morphology and poor proliferation of 3T3-J2 cells after a few passages is often indicative of cellular stress, senescence, or suboptimal culture conditions. Although cells may initially appear healthy post-thaw, stress responses can manifest after subsequent passages. Possible causes include high passage number, which is known to affect 3T3-J2 phenotype and growth potential, or serum quality; variability between FBS batches can significantly influence attachment and proliferation.
Also, consider that trypsinization conditions, overexposure, can damage cells and affect viability. Ensure that media components are fresh, pH and CO₂ are well-controlled, and that cells are not over-confluent or seeded too sparsely. If these factors are optimized and the issue persists, it is advisable to obtain a fresh, low-passage vial from a reliable source.
Maisra Azhar Thank you for your answer. My colleagues use the same media and FBS batches for their cancer cell lines, and they work perfectly fine. Is this sensitivity specific to 3T3-J2 cells?
Also, I noticed that some papers used BCS instead of FBS. Does this also affect cell growth?
Regarding BCS (bovine calf serum), it contains a different composition of growth factors and proteins compared to FBS, generally being lower in mitogenic activity. While some protocols report successful use of BCS with 3T3-J2, it can significantly affect proliferation rates and morphology, and should only be used if validated for your specific application. If you are at the stage of protocol optimization, then you can use it to check and develop a protocol.
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