http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2326106/

I use TE-7.

Vimentin is the most commonly used

I use vimentin primary antibody to identify fibroblast cells.

Anti-vimentin antibody is good in your request.

There is no single reliable immunostain for fibroblasts. The safest approach is to incorporate H&E appearances,the positive reaction to Vimentin and the negative immunoreaction with epithelial/ mesenchymal cells to make a reliable interpretation.

Agree with Ali Hussein. I am a histopathologist. The identification of a fibroblast is inaccurate. We do not know the identity of all cells that look like fibroblasts. Some may be mesenchymal stem cells (for example ALDH1+). The identification of fibroblasts is (apart from Vimentin positivity) a matter of exclusion. We can use markers for contractile proteins, such as alpha smooth muscle actin (SMA), and call fibroblast-like SMA+ cells "myofibroblasts". Also, myofibroblasts are much less positive to smooth muscle myosin heavy chain than they are to SMA. This issue is far from fully elucidated. Thus exciting.

Vimentin would be my first choice as well. Together with a spindle-shaped morphology strong staining should be a pretty reliable and most stable marker for fibroblasts, though quite a few other cell types will show weaker or variable staining, too.

Thanks all for the fast feedback, I very appreciate the discussion.

I was a bit reluctant to use vimentin since it seems to stain lots of other cells too (http://www.proteinatlas.org/ENSG00000026025/tissue/tonsil#img) but will give it a try...

keep up the good work !

Vimentin is expressed in mesenchymal cells - it's not per se a fibroblast marker. It's widely used as a fibroblast marker though - from the histology and the spindle like morphology of the fibroblasts they should be easy to identify. Clone VIM 3B4 would be my suggestion.

You can also try a combination with smooth muscle actin and/ or for skin fibroblasts Heat shock protein 47 (HSP47).

Vimentin is the best marker for fibroblast. Other markers are: anti-alpha-actin clone 1A4, anti-CD34, anti-HSP47 (Heat Shock Protein 47), anti-SFA (Fibroblast Surface Antigen). It is necessary to do some preliminary test to choose the best one.

Axel, I am afraid that I have to agree with the rest of the people. There is not one fibroblast -specific antibody as far as I know. Vimentin is a good candidate but it can stain other types of cells too. I think that you need to try a few of the options suggested above especially Seppe's idea with FSP-1.

hi axel,

You may be aware that there is no single fb marker that will demonstrate all fb in a given synovial section; the closest to this would be prolyl-4-hydroxylase whihc will staing lining layer and sublining fibroblasts but you would have overlap with endothelial cells. Similarly CD90,Thy1 will overlap with EC but only stain sublining fb! I think you need to refine your question in terms of synovial geography; do you wish to see both LL and SL fibroblasts, or only SL fb that might interact with plasma cells, for instance? The second question would eb how many colours/stains are you limited to? Because of the complexities of staining stromal cells it's dificult to help further without more detail!

I usually use CD34... good luck!

Yes, specifically marking fibroblasts is a hard task... if you're looking for activated fibroblasts/myofibroblasts I would go primarily with alpha smooth muscle actin, Hsp47 (collagen production). These two should not be in the inflammatory compartment... FSP-1 sounds great, but I never had luck with it. Vimentin is generally present in connective tissue cells, including blood/inflamatory cells.

Axel - don't lose heart - I agree that vimentin is actually not very specific (high sensitivity but low specificity) and also that FSP-1 is probably inappropriately named - we also cannot reproduce useful staining with this. SMA is unfortunately also only sparingly expressed in the synovium. But if you can specify which part of the synovium you wish to stain then there are likely to be markers that will help you. Fors instance in the sublining you could use CD248, while in the lining layer you can use VCAM-1, CD55 or even cadherin-11. I would still encourage you to stain for one or other of the subunits of prolyl-4-hydroxylase (collagen synthesis) - if you are using immunohistochemistry then excluding endothelial staining should not be difficult.

Dear Axel,

A few antibodies have been previously reported to recognize fibroblasts, some of which take advantage of fibroblasts as the major producer of collagen. One monoclonal antibody (anti-pC) is directed against the cleaved carboxy terminal propeptide of the triple-helical procollagen molecule. This antibody stains some of the fibroblasts from patients with active pulmonary fibrosis, but not fibroblasts that are not actively dividing . 1B10, an IgM antibody that reacts with a surface membrane protein, has also been reported to be fibroblast-specific. The 1B10 antibody is reported to recognize human fibroblasts, tissue macrophages, and peripheral monocytes. Yet another reported anti-fibroblast antibody, 5B5, is directed against prolyl-4-hydroxylase. This enzyme catalyzes a hydroxylation reaction that is a prerequisite for triple helix formation of the collagen protein. The 5B5 antibody recognizes fibroblasts in loosely cellular regions, but false-negative staining was observed for resting, mature fibroblasts in dense connective tissue.

In formalin-fixed cell pellets, the TE-7 antibody is more specific than the 1B10 or 5B5 antibodies . The TE-7 antibody recognize both dermal and lung fibroblasts and both growing and quiescent fibroblasts. The 1B10 and 5B5 antibodies are less specific; they recognized fibroblasts, but also recognized the non-fibroblast cell types.

Hi All,

Have any of use used the TE-7 Antibody to stain fibroblasts in mouse heart tissue?

And the morphology of the cells in your section should also be observed. SOmetimes a good H & E is a good "special" stain. The saffron compoment (in HES slides) can also be of use. Otherwise - CD34 or vimentin; and no single stain will necessarily tell all the tales.

Hi Axel, together with an elongated, spindle-shaped morphology strong vimentin staining should be pretty reliable to identify bona fide fibroblasts. Certainly, vimentin is present in many other cell types, though at lower levels, and can be induced transiently even in epithelial cells.

(My previous answer had disappeared from the screen)

Hi Dr Ashraf Imam. What you say about using Vimentin and Alpha-Smooth Muscle Actin is very interesting and will be very useful. Can I trouble you for references to support this. Thank you and kind regards.

Just my two cents... Vimentin is the intermediate filament in mesenchymal-type cells, thus all connective tissue cell types will express it - unfortunately the leukocytes themselves are vimentin positive too. Of course, there are many vimentin antibodies, I can not exclude that there are some which shows more affinity to fibroblast than leukocytes. In context with inflammation, wound healing therefore I do not use vimentin. Instead I use anti-Hsp47, anti-pro-collagen and alpha-smooth muscle actin. These antibodies will pick up the fibroblasts and myofibroblasts characteristic to inflammatory environments.

I use vimentin to stain fibroblasts

It seems like many types of epithelial cells also become vimentin positive in vitro:Kasper, M. and P. Stosiek, The expression of vimentin in epithelial cells from human nasal mucosa. Eur Arch Otorhinolaryngol, 1990. 248(1): p. 53-6.

Mendez, M.G., S. Kojima, and R.D. Goldman, Vimentin induces changes in cell shape, motility, and adhesion during the epithelial to mesenchymal transition. FASEB J, 2010. 24(6): p. 1838-51.

Rogel, M.R., et al., Vimentin is sufficient and required for wound repair and remodeling in alveolar epithelial cells. FASEB J, 2011. 25(11): p. 3873-83.

FR. PIEPER, FA. VAN DE KLUNDEKT,Regulation of vimentin expression in cultured epithelial cells Eur. J. Bioctiem. 210, 509-519 (1992)

Vimentin is your best bet.

Caution: In HE stained tissue sections of normal stroma of human mammary and urinary bladder, many of the fibroblast-like cells are stellate cells (a k a vitamin A-storing cells / Ito cells). Can be confirmed by IHC for either cellular bindig protein 1 (CRBP-1), vinculin (although also endothelial cells are positive), or aldehyde dehydrogenase 1 A1 (althhough some round/oval mast- or MSC-like cells are positive).

Hi,

I am staning for fibroblasts with TE-7 and was wondering if there is any protocol for staining in paraffin embedded tissue for this type of antibody?

I tried vimentin but it did not really.

Article An Immunohistochemical Method for Identifying Fibroblasts in...