I have just isolated some tail tip fibroblasts from very young (1-wk old) mice for experimentation. I am using IHC to confirm the identity of the fibroblasts.
I have looked at a few Q&A topics on ResearchGate and also some papers for some good surface markers. So are there any other markers besides Vimentin, FSP, SFA, TE-7, CD34, that generally stain fibroblasts (with a lot of exceptions), that can be more specific to identify my cells as Tail Tip Fibroblasts.
Also I am so confused about CD34, isn't it staining stem cells? Or tail tip fibroblasts are multipotent? Okay I am really confused now...
Thank you for any advice or suggestions.
PDGFR-alpha is a marker of fibroblasts; CD34 is supoposed to stain interstitial fibroblasts in human tissue. CD34 staining is a mark of undifferenciated fibroblasts meaning they are stem cells. The suffix "blast" is used in cellular biology to denote a stem cell or a cell in an activated state of metabolism. fibroblasts play a critical role in wound healing meaning that they regenerate damaged tissues which is a characteristic of a stem cell...
Other twoi important markers for fibroblast are: SMA (smooth muscle actin), TCF-4 and CD90. SMA is not very specific but fibroblast must show high SMA expression. TCF-4 and CD90 are more fibroblast specific markers.
Hello
Fibroblasts are cells that form the connective tissue. They can differenciate into distinct connective tissue cells and maintain the extracellular matrix of the fibrous connective tissue.
for surface markers you have a good choice , and I'll explain to you :
Vimentin ; is the most frequently found intermediate filament in fibroblasts .
SFA : is a glycoprotein produced by connective tissue cells e.g (mesenchymal cells, fibroblasts, and astroglia cells) that is also suited for detection of fibroblasts and fibroblast cell lines.
for antigen CD34 is a good choice and i am agree with Célimène Galiger ,where there is in human hematopoietic progenitor cells , It is distributed in most connective tissues, among them alveolar connective tissue and the connective tissue of skin and blood vessels. A marker for fibroblast differentiation in the human dermis are endogenous peroxidases it can also be used to mark skin fibroblasts
Fibroblast-specific protein 1 (FSP1) can be detected in mice as early as day 8,5 p.c. It occurs in the cytoplasm of fibroblasts, but not in epithelial cells. Rarely, FSP1 antibodies will react to the parenchym. In case of inflammation, epithelial cells are marked by FSP1 antibodies, too, which are thus assumed to be converting to fibroblasts at the site of inflammation.
For activated fibroblasts, we recommend procollagen type 1 or Prolyl-4-Hydroxylase beta antibodies, the latter occurring in epithelia, endothelia, alveolar macrophages and in interstitial cells.
To detect myofibroblasts, the anti-alpha-actin antibody with the clone designation 1A4 from smooth muscle tissue can be used.
They express ER-TR7, intracellular components which are thus very practical to use for marking such fibroblast types.
Ref : (Strutz ,F. et al. 2004 ; K. Kuroda und S. Tajima, 2004)
Best wishes
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