Hi all,
We are growing the non-cancerous human breast cell line HMT3522 S2 using the media CM, including:
- 500 ml DMEM/Ham’s F12 (Gibco, 11039-021),
- 1% Insulin-Transferrin-Na Selenite,
- 10 ng/ml EGF,
- 50 nM progesterone,
- 1% Pen/Strep (Sigma, P3539).
However, they grow very slow, cannot attach to the flask, and seem not to be healthy.
Could you please give us some advises to grow them.
Thank you in advanced.
DMEM / Ham's F12 (1:1) + 2mM Glutamine + 250ng/ml insulin + 10µg / ml transferrin +10E-8M Sodium selenite + 10E-10M 17 beta-estradiol + 0.5µg/ml hydrocortisone + 5µg / ml ovine prolactin
some ingredients seem to be missing
* If in doubt, add FCS
* If still in doubt add appropriate nonionic detergent or polymer carrier (research options)
* If still in doubt remove pen/strep as it is often a problem for cell lines (yes, you have to have impeccable sterile technique)
DO test for Mycoplasma. Half of all shared cultures are infected.
Thank you a lot, Dr Artem.
Do you have to grown them on a collagen-coated flask? If yes, which one you have used, collagen type I or IV?
I read some documents that mentioned this cell line should be grown on a serum-free medium. Why FCS can solve problem?
Many thanks
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