Hi Ruhul,

Good news.   As part of the Breast Cancer Campaign Tissue and Cell Bank initiative I isolate normal human breast epithelial, myoepithelial and fibroblast cells.  These primary cells grow in culture  in appropriate media for a number of passages and stay pure.  If you're interested in applying for these cells, or indeed cells from other types of human breast tissue, please look us up on the web.  We will also provide you all the help you need in your experiments.  Look forward to hearing from you.

Thanks Sylvia, I think HMEC has basal characteristics. I was looking for normal cells that has typical luminal characteristics.

http://www.lgcstandards-atcc.org/products/all/CRL-10782.aspx?geo_country=gb#characteristics

what about MCF12?

Hi, Sylvia Riedel, Ruhul Amin is talking about normal Human mammary luminal cells not Breast cancer cell. Remember he specifically said Normal

I would also like to know if anyone has used any commercially available normal human breast cell lines

I used to use MCF12 cells, which are cmmercially available. Velocity of growth is quite slow compared to cancer cell lines, but it is OK

Thanks Kenji. MCF12A can be an option. But according to ATCC, it's K14 positive which usually is a basal marker. Do you think it will be ok to use it as a normal differentiated luminal cell?

ATCC says MCF12 cells are  typical luminal epitherial breast cells. If CK14 positivity is your concern, MCF10 could be an alternative choice. 

Sorry Kenji, MCF10A is more basal than MCF12. Luminal cells should be CK18/CK19 positive and low CK14.

I would then suggest you to check this website. Hope it helps you out!

http://www.atcc.org/~/media/PDFs/Cancer%20and%20Normal%20cell%20lines%20tables/Breast%20cancer%20and%20normal%20cell%20lines.ashx

Thanks Kenji. Let me check.

MCF10A cells are from a non-malignant hyperplasia (from a fibrocystic disease) and is often used as  a 'normal' control. Yes, they do express CK14, but they are not myoepithelial (if that's what you mean by 'basal'). Primary epithelial cells in vitro are CK18/CK14 double-positive (as are MCF10A). Even the non-malignant mouse mammary epithelial cell lines EpH4(K6) and HC-11 are CK14/CK18 pos.

There are plenty of companies selling HMECs (which are also CK14 pos.). I believe there is a Tert-immortalised cell line around.

Thank you Dr. Torsten  for the clarification. Actually I was looking for something like MCF7 but normal. My MCF7 cells strongly express CK18 but low CK14. If  that is the case for CK14, then I can use  MCF12A as Kenji  suggested. What do you think about that?

MCF12A will be very much the same as MCF10A in this  respect. They both express basal-cell cytokeratins.

It's hard to say what the best control is for you, as I don't know what you want to use it for. Neither MCF10A nor MCF12A is a normal differentiated luminal cell line - differentiated cells would not really grow in culture as they are differentiated. I don't know of a 'normal' (or better non-malignant) cell line that has the same hormone receptor and CK expression status as MCF7 cells. By the way, MCF7 cells can also be highly variable from lab to lab as it's genome is pretty much messed up. Depending on your question, it might even be better to compare against other BC cell lines. In any case, you should not rely on just one cell line. 

Sorry if this doesn't really help you.

By the way, you may want to look at this recent paper. I haven't properly read it myself yet, but it might be helpful.

Sauder et al. BMC Cell Biology 2014, 15:20

http://www.biomedcentral.com/1471-2121/15/20

Phenotypic plasticity in normal breast derived epithelial cells

Thanks Dr. Torsten for your kind reply. I was also confused regarding the use of MCF12A. One option might be the use of sorted cells from reduction mammoplasty or any commerical source. Actually I was comparing the differential expression of some genes among basal (normal/cancer) and luminal (normal/cancer) lineage. In this regard, I have already investigated my hypothesis on MCF10A, MB-231, Hs578T, MCF7 (Luminal A), T47D (Luminal B). Can you suggest me some other luminal A/B type BC cell lines?

How about SK-BR-3 or BT474?

Check out this paper from Mina Bissell's lab. This will give you a few options.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2391005/pdf/nihms34890.pdf

Good luck!

I have not come across any cell line that has similar features and functions as that of 'normal' primary human mammary luminal epithelial cells that is produced in vivo. If you want something that 'kind-of' works like luminal then go ahead and try MCF10A. Its not perfect cell line but can get you going depending on your questions.

Another option is to buy HMEC cell line which contains mix of basal and luminal. You may be able to FACS or immunomagnetically purify the luminal fraction from the mix and culture them separately.

Last option is that you find a lab that routinely isolates 'normal' primary cells from breast tissues and request them to provide you with luminal fraction. Best wishes. 

Thanks Dr. Torsten, usually SKBR3 is mostly studied as Her2 overexpressing cells, right?  The status of many cell lines are confusing since many researchers categorize them differently.

Thanks Dr. Nagarajan, I used MCF10A but I found it basal-like. I was also thinking about FACS sorting using CD49f/EpCAM though. How about inducing luminal differentiation in  MEC by matrigel/prolactin?

You could try sorting them, but I fear that if they grow in culture they will eventually take on a CK14/CK18 double positive phenotype.

Yes, it is confusing, probably because it's not so clear cut, especially with cell lines or cells in culture. However, check out the WB description of the different cell lines in the Kenny et al. paper. That might help you.

Hi Ruhul,

Good news.   As part of the Breast Cancer Campaign Tissue and Cell Bank initiative I isolate normal human breast epithelial, myoepithelial and fibroblast cells.  These primary cells grow in culture  in appropriate media for a number of passages and stay pure.  If you're interested in applying for these cells, or indeed cells from other types of human breast tissue, please look us up on the web.  We will also provide you all the help you need in your experiments.  Look forward to hearing from you.

Hi Jenny,

This is really good to know. Do you by any chance distinguish between intra- and interlobular fibroblasts as well?

For how long do the luminal cells retain their CK expression profile? I guess these will be mostly ER neg?

Hi Torsten

We do isolate fibroblasts by several different methods.  Some would favour intra and some inter fibroblasts.  We haven't distinguished between them ourselves.  Perhaps you would like to give it a go.  We also isolate them from a variety of tissues and have tumour and surround tissue sets available. The primary  luminal epithelial cells survive in culture for up to 3 passages but the primary myoepithelial cells will keep going for 6 passages.  They both retain their CK8 and CK18 profile for epithelial cells and CK14 for the myoepithelial cells.  Both types of cells are in sufficient numbers to do a whole variety of experiments. We have many years of experience using them ourselves!   We have also recently been able to culture luminal epithelial cells with ER expression which is quite an achievement in itself but, because they are normal cells, it is only a small percentage.  The cell bank part of the BCC Tissue bank is relatively new and we are just starting to distribute.  But we have amassed a considerable repertoire.  So come up with a project and we will help you fulfil it.

Hi Jenny,

This sounds great. I will be in touch soon if that's OK. I am especially interested in the fibroblasts (CAFs and NAFs), but also the primary luminal cells. 

BTW,  I am actually BCC funded, and these could help me in my current project  as well.

That sounds perfect.

Email me at work on [email protected] and we can chat about how we can help you.  Once you've decided exactly what you want there is an official application process. But if you've already got past me it's more or less a formality.  There are also costs involved but they are minimal and as you are BCC funded it should be fine.  Looking forward to hearing more about your project.

Thank you very much Dr. Jenny for your great news. I am really interested in applying for the cells. Could you please tell me how the application process works? And also about the expenses outside UK?

I am using a cell line that we ordered from Sigma, called 1-7HB2:

http://www.sigmaaldrich.com/catalog/product/sigma/10081201?lang=de®ion=DE

They have cuboidal morphology and behave pretty much as luminal epithelial cells. I can't say what is the expression pattern with regards to basal/luminal cytokeratin markers though.

I hope this can help.

I prefer Primary culture from human origin