i am growing MDA MB-468 cells in my lab. For few weeks we grow them in L-15 media but they were growing very slow , it took almost 20 days to become 25cm flask to get confluent . Now we shifted them to RPMI media as suggested by some experts but still after 2 weeks they are not growing well and there are many small dot like structures appearing on flask. how can I get rid of these dots and get a normal proliferation rate in cells?
hi Heena,
You sould complete your media (RPMI or DMEM) with S/P, 10%FBS, 1mM L-glutamine (if the media does not contain it), 1x Non essential amino acids, NEAA (optional), 1 mM sodium pyruvate (extra carbon source, optional). Sometimes some breast cancer cell lines need extra estrogen for their growth. Please check if your cell line needs estrogen or not?
I hope this helps
Hello
These cells require L15 medium (Leibovitz medium), with 2mM L-Glu, 10% FBS and grow in incubator without CO2 . perhaps damage to the cells at freezing and thawing might slow .
for RPMI : yes you can try with it + 10% FBS .
RPMI was a better medium than DMEM and for these cells.
https://physics.cancer.gov/docs/bioresource/breast/NCI-PBCF-HTB132_MDA-MB-468_v1.0_SOP-508.pdf
rpmi media or l15 are not critical most probably the cells can grow either in both conditions
but the co2 free environment is very crucial for them ! !
I'm pretty sure those small dots are dead cells, but check for mycoplasma to be sure(it's a good idea to check your cells at least once a year anyways). It could be just a bad freeze so getting a new vial from the vendor would be a good idea. Also make sure your incubator is functioning properly(temp and CO2 levels are accurate).
The suggested culture conditions and reference are below.
-Dulbecco’s modified Eagle’s medium/F-12 supplemented with 10% fetal bovine serum (Gibco, Carlsbad, CA) and antibiotics in a humidified atmosphere of 5% CO2.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4065802/
Mycoplasma is the main cause of cells growing slowly. Check mycoplasma and if positive, there are many successful treatments.
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