I want to identify the mast cell by flow cytometry from mouse peritoneum.
Mast cells play an active role in variable diseases such as atherosclerosis, asthma, arthritis, bile duct fibrosis, pulmonary fibrosis, and malignant tissues characterized by melanoma. Trypsin-like neutral serine proteases are well-known to be predominantly expressed in mast cells. Furthermore, CD203, which belongs to ecto-nucleotide pyrophosphatase/phosphodiesterase（E-NPP）family proteins, highly express in both mast cells and basophils. That is why the IHC staining with the combination of trypsin-like neutral serine proteases and CD203 is useful to accurately identify mast cells. In the case of FACS analysis, the cell surface marker CD203 is the important molecule to purify the population of mast cells.
CD203c is also expressed by other cells (e.g. basophils), so I would not recommend using only CD203c. Intracellular tryptase is best, but if you want to use extracellular markers you could use the combination of CD117 and FcERI.
Thanks Yoshida and Jolien,
As mostly I need to identify the MC for the in vivo work so I think extracellular markers will save me time and convenient to use. I will use CD117 and FcERI.
Thanks both of you
HI >> DOCTORS
MY NAME IS OMAR AND I MASTER STUDENT IN ORAL PATHOLOGY
I NEED TO HELP ME IN SOME PINTS IN THIS TOPIC
MY RESEARCH IN role of TLRS IN MAST CELL IN SQUAMOUS CELL CARCINOMA
SO >> IN MOUSE
SO MY QUESTIONS ?1- HOW I CAN ISOLATED MAST CELL FROM ORAL MUCOSA( BEST MARKER)
2 - BEST TO ISOLATED TLRs on mast cell?
3- there is way to help me in make steps (in question 1,and 2) in one step >>> for economic reasons ?
thanks
Hi All,
Can anyone advise about good antibodies to use to detect murine mast cells in FFPE tissue (lungs and skin). I normally use mast cell tryptase for human tissue but I can't find that for mouse.
Hi Peter,
If your goal is it just detect the murine MC in lungs and skin tissue, you can use the Toluidine blue staining, We regularly use and MC are very clearly visible and you can identify the activated (degranulated) MC also.
Good luck
Thanks Roopesh,
I have tried toluidine blue with tryptase in human lung, it only identifies about 60-70% of the mast cells. Is it better in mouse?
I do not have experience with human samples, but in mice samples we can clearly see all the MC, the staining is very nice and if you wash a bit more then the background noise is no more. We use acidified TB. If you a sample to try, I would suggest you try with your hand.
Roopesh
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