Has anyone here experience with culturing Breast cancer stem cells (Parental)?

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Utsav Sen

Hi Jamail,

I am using breast cancer stem cells, and culturing with 10% FBS media supplemented with 1%Penstrep,1%L-glutamine,1% non-essential amino acids, 1% of sodium pyruvates and 0.1% of beta mercaptoethanol

Try this media; it will work fine.

All the best!!

Utsav

Ming Chuan Yu

Hi Utsav Sen. My intention is to get more pure stem cells and keep them in high stemness. I do not want them to be differentiated. It seems that you are culturing to differentiate as you add 10% FBS and in regular CO2 incubator.

I am culturing them in hypoxic incubator with 1% FBS or Serum free. Where did you get your BCSCs? Is that Parental or Pure BCSC?

I bought BCSC from Celprogen but Celprogen said BCSC (Parental) include 2 types of cells and I have to select to get pure BCSCs which are rounded cells but not spindle shape elongated fibroblast-like cells. I am not quite sure what Celprogen was saying. Also cannot find any protocol in their website and published papers.

My intention is to run MTT test with BCSCs. Since my cells are not Pure BCSCs, I am not sure how to run MTT with them.

I have sorted the breast cancer stem cells from wild-type population. I have used CD44+/24- surface marker to isolate the cancer stem cells. BCSC were isolated from commercially available MDAMB 231 or 468 cell lines. Their stem ness and other molecular characteristics are not going down.

Please, see this paper, where I have used colon cancer stem cells and same media. You will find the stem ness phenotype, and the other characteristics are present in those cells. https://www.ncbi.nlm.nih.gov/pubmed/28755485

Whichever cell line you bought from the company, follow the protocol that has been provided by them. But you can try the above mentioned media for one experiment.

Pure BCSC will have mesenchymal morphology as I have seen, and it's reported in literature too.

Regards,

Thank UtSav. I am now a bit confused for adding 10% FBS and using regular CO2 incubator because I notice to reduce FBS or make FBS free and to use only 5% O2 rather than 20% O2 when we are culturing Cancer Stem cells or stem cells. What is your opinion or experience over these.

I did try usign 10% FBS for cancer stem cells, all cells become fibroblast-like cells and growing fast. So, as per your suggestion above, 10% FBS does not give differentiation to BCSCs right? I do not know why others saying must reduce serum and also must use hypoxic incubator.

Thanks for your paper link. Will download and read.