I am generating astrocytes from stem cells and want to prove that they are actually postmitotic and not only quiescent precursor cells. Are there any reliable markers except for incorporation of nucleotide analogues during DNA synthesis?
Nucleotide analogues, Ki67 or PCNA would label actively dividing cells, and as you want to also exclude quiescent stem cells you can't use GFAP. How about using S100Beta? According to Bonaguidi et al. 2012 and Kronenberg et al. 2007 it labels (edit: only) postmitotic astrocytes.
If you dont have any differentiation marker specific for your cell-type why not look for absence of proliferation markers (phospho-Histone H3, Ki-67 etc).
Thanks a lot for these fast responses! The cells express S100b, but also the precursor cells do. I know that the cells express markers of 'mature astrocytes', but still I want to see if they are really postmitotic, since this is not so easy to define for astrocytes. Of course, I can confirm it by the absence of proliferation markers, but I am not happy with this. For me, it's better to show that something is present. I was thinking about trying to stain p21 or p27. What do you think?
Ok but by staining p21 / p27 you still could not rule out quiescent progenitors. Depending on your specific type of progenitor, you could co-stain for Nestin - differentiated astrocytes should by negative, while quiescent stem cells express it (Song et al. 2012).
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