Hi, I bought 3T3-L1 cells from ATCC and would like to differentiate them into white adipocyte-like phenotypes. I thawed the cells and cultured them in DMEM+10%BCS+1%P/S (37C, 5%CO2). The medium was freshly prepared before thawing. Starting from the third passage, however, many cells are in suspension and only a few are adhesive to the tissue culture flask. Both pictures and videos are below. Has anyone had experience with this cell line and what is wrong with my protocol? Any suggestions would be greatly appreciated. Thanks!
Hello Yichen,
Is this the medium that ATCC recommends? This is critical; otherwise, ATCC will not reimburse you if needed.
Have you tested the medium for mycoplasma?
The culture looks very crowded. Have you tried seeding the plate with a lower cell concentration?
Michael Firer
Thank you michael for the suggestion.
We don't have mycoplasma test kits right now but I will consider buying one.
I changed the cell medium the day before and at that time the cell confluency is relatively low (20%), and the cells are still bearing pre-adipocyte phenotype. In this case I don't think it's caused by high cell concentration, right? Please let me know if you have more suggestions!
I may be wrong but the videos look like it could be a fungal (yeast ?) contamination : has it gone away after freezing or persisting?
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