What are the best markers for Erythropoiesis in mice?
There are several markers like CD45, CD34, CD117, CD105, CD235a, CD71 based on specific stages of erythropoietic cells.
Which of the above or novel markers or a combination works best to recognize the erythropoietic cells?
Also, is there any better method to assess the rate of erythropoiesis in mice?
The best way to asses eritropoyesis is an RBC count. You do not need sofisticated methods for that.
Dear Sandeep Rai Kaushik ,
Murine hematopoietic progenitor cells (HPCs) express different markers compared to human HPCs. For example, they are not characterized as CD34+. CD34 is usually used for identification of common myeloid progenitors (CMPs) and granuclocyte-monocyte progenitors (GMPs).
By the way, for the flow cytometric analysis of erythroid progenitors and precursors, I would like to recommend staining of the following markers.
(Minimum requirements) Kit (CD117), Sca-1, CD71, Ter119
(Additional markers) CD34, CD31, CD16/32, CD41
First of all, undifferentiated erythroid progenitors are found in the Kit+Sca1- fraction. In that fraction, you could enrich burst-forming erythroid (BFU-E) progenitors and colony-forming erythroid (CFU-E) progenitors by gating CD71(low) and (CD71(high) cells, respectively. Those erythroid progenitors are usually Ter119(low).
If you are interested in more differentiated erythroid precursors and erythroblasts, then you may find those cells by gating Kit-CD71+ cells. Those erythroid cells are usually Ter119(high). Thus, the Ter119 in mice can be used in a similar manner to the CD235a in humans.
Hope this could help.
Thank you Tetsuo Tsukamoto for the detailed explanation.
I am still curious to understand which population will be a better indication of erythropoiesis rate (kit+sca1- vs kit-CD71+), undifferentiated vs matured/more differentiated.
Thanks, Tomas Koltai .
Can you please explain how measuring the RBC count will tell the erythropoiesis rate?
Please note that I am coming from another field and not yet very experienced in analyzing erythropoiesis, so I do not have a definite answer. Nonetheless, I suppose it might depend on the environment your mice are placed. As you know, anemia can be caused by several reasons including the lack of iron, the lack of progenitors, the lack of vitamin B12, etc. The rate of erythropoiesis may be regulated by different factors depending on the setting. So, I suppose you could clarify the factor of your interest. Are you interested in BFU-Es, CFU-Es, erythroblasts, reticulocytes, or whatever? Any one of these could be the regulator of the rate of erythropoiesis, I just suppose.
Thank you Dr Tetsuo Tsukamoto .
I am not interested in a specific population but in knowing the rete of erythropoiesis by the best available option. So, I am flexible with the cell population or even the technique.
I suppose that in the steady state erythropoiesis, all of those I listed are involved in defining the homeostatic rate of erythropoiesis. And you would eventually need to determine yourself which one(s) to have a close look at, I just suppose.
For details on the actual methods for analyzing the rate of erythropoiesis, I could recommend to read better references such as the following (that is expensive, though).
"Erythropoiesis - Methods and Protocols"
https://www.springer.com/jp/book/9781493974276
I could imagine methods such as in vivo labeling of all RBCs followed by tracking of unlabeled (newly generated) RBCs (no idea how well such a method will work), and in vitro differentiation (enucleation) of BFU-E or CFU-E cells (needs FACS sorting of those cells and cytokine-supplemented medium). I have not tested any assay myself yet.
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