Hello everybody,
I am going to cultivate MDA-MB-231 cell line along with three other cell lines. I want to know if I can cultivate this cell line with RPMI-1640 Medium under the following conditions: air, 95%; carbon dioxide (CO2), 5%? Does anyone have experience with this?
(Since culture conditions of Other cell lines are RPMI-1640 Medium, air, 95%; carbon dioxide (CO2), 5%.
Thanks in advance.
I agree with Daniela. DMEM is quite a standard and highly recommended medium for growing MDA-MB-231 cells. ATCC uses complete L-15 (+10% FBS). Most papers talk about culturing MDA-MB-231 cells in DMEM. Some resources are attached, I have highlighted the areas need to be focused.
Good luck!
The question is not in my area of expertise, I suggest you direct them to a biologist rather than a geographer
Dear Katarzyna Julia Marosz,
Thanks for your suggestion, but as far as I know, I did not ask a geographer. but again thank you.
5% CO2, 37°C in a humified incubator is ok, but I always used DMEM/F12 medium, DMEM medium is suggested.
I agree with Daniela. DMEM is quite a standard and highly recommended medium for growing MDA-MB-231 cells. ATCC uses complete L-15 (+10% FBS). Most papers talk about culturing MDA-MB-231 cells in DMEM. Some resources are attached, I have highlighted the areas need to be focused.
Good luck!
Hello, I use high glucose (25mM), 1mM sodium pyruvate DMEM (1995-065 from ThermoFisher) with 10% FBS and 1% Pen/Strep to culture my MDA-MB231 cells, although RPMI would work as well.
Dear Arpan De,
Thank you for your answer and attaching Some resources.
What I would suggest, if someone in your group has worked with these cells before get their opinion first. Use a protocol that was used in your lab to cultivate these cells because as you can see many people have different was of culture them and their all going to be a little different. But if they are new and its your first time, I always liked to consult ATCC. Good luck with your cells.
https://www.atcc.org/Products/All/HTB-26.aspx#generalinformation
Dear Reyhaneh,
Yes you can, we are using the same conditions RPMI 1640 with 10% FBS @ 37 degree, 5% CO2
MDA-MB-231 breast cancer cells cultured in different types of media (MEM), (DMEM), or (RPMI)-1640 medium] containing different concentrations of fetal bovine serum (FBS) or different sera that were grown at different cell densities.
Dear Miss Ravanbakhsh,
As I had similar problem, I only can suggest that culture these cells by using DMEM low glucose, When your cell initiate to growing in exponential phase, subculture them in the different culture flask and use RPMI 1640 with 30-40% FCS.
Plee remember that the exponential phase is the most important point for changing the media.
Dear Abolfazl nosrati tirkani, I don't think that 30-40% FCS is anything good for the cells, appart from being expensive cell culture. There is no sense to go for this high FCS content unless you want to create very FCS-sensitive cells which will definitely have a very different behaviour than any normal MDA-MB-231 cells grown under 10% FCS. I have worked with plenty different cell lines from various sources, but we never used more than maximum 20% FCS in very rare cases or for thawing of very sensitive cells for the first 1-2 days. With 30-40% FCS content you will not get any reliable results from experiments as extensive amounts of growth factors, cytokines and other factors from the FCS will make your cells behave different than under normal conditions using 5-10% FCS
.
Thank you for your recommendation, you know I meant when sb wanna to change the entire cell culture condition, he/she has altere some established rules. I know thode amount of FCS is not appropriate for any type of cells but as I said, it will be change under different condition as such changing the media,
Anyway thank you very much for your suggestion,
I just explain my experience!
Thanks to all contributors.
Yes, in my 9 years experience of working with the MDA-MB-231, along with other ER+ve/-ve BC, as well as other cell lines, i found a mixture of 1:1 ratio of RPMI-1640:DMEM media to give the best conditions for these cells.
Most of the times these cells did attain 90-100% confluency in just 48 hours, using only 5% FBS, which still falls within the product guidelines provided by ATCC, the original, lower-passage suppliers of the MDA-MB-231 cell line.
In fact, for all the nearly 30 differents cell lines (both cancer & transformed-normal cells) i happened to culture, propagated, stocked-up and experimented with, i have in deed NEVER EVER had to use anything more than 15% FBS/FCS.
The only exception here was on temporary (not for continuous) cell culture, involving whole-blood-based Human Lymphocytes, for cytogenetics-karyotyping/genotoxicity studies, where i had to use 20% FBS.
In conclusion, i would like to suggest that, in order to have optimal growth conditions while saving costs simultaneously, please go ahead Reyhaneh. Set up your culture using a separately-aliquoted with 5% FBS supplemented medium for MDA-MB-231 only (or, RPMI-1640 with 10% FBS), at the standard culture conditions of 5% CO2, 95% Air & 100% humidity.
Hello, it's nice to see these excellent suggestions. I have a question for the culturing of MDA-MV-231. If I use complete L-15 (+10% FBS), can I still cultivate the cells with 5% CO2? Because I saw that ATCC recommend the condition without CO2 but it's more convenient for me to culture this cell with 5% CO2. Thanks so much.
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