Etchevers:Notebook/Genomics of hNCC/2008/06/23
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Rewrite. Future directions.
I was rewriting for the n'th time our manuscript at the request of the current journal editor, who would like us to turn the focus back towards stem cells. We know we'll be shot down for not having the functional data and being too descriptive yet again. But it is worth anticipating what we will do.
I have requested some serum replacements for testing on the hNCC lines I have here in Toulouse: R1064 and the barely surviving R1094. I will have to ask for new cells to be sent to me in August. (Although I am hardly present in August either, but I think I can propagate cells.)
Had devised - at least partially - a qPCR assay. But to get stat sig results need 6 lines and 6 control lines (thinking - with and without this serum lot) and that will never happen as I am running out of serum and am not sure that it still has the original activity after this last thaw-aliquot-freeze. So will have to just test the serum adjuvants blind and see which reproduce the best propagation, and perhaps a couple of immuno markers (NCAM-SMA-DAPI profile is pretty easy to recognize).
Once that is established, can move on to in vitro differentiation.
To propagate, need to inhibit phospho-ERK as well as GSK3 (activated beta-catenin) according to Ying/Smith May Nature paper.