Etchevers:Notebook/Genomics of hNCC/2008/06/23

From OpenWetWare

Jump to: navigation, search
Genomics of human neural crest cells Main project page
Previous entry      Next entry

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.

  • Sophie had contacted someone in M. Cavazzana-Calvo's group for eventual blastocyst injections - for which we certainly need to request ethical approval - but she is out on maternity leave now so it will have to wait until the autumn.
  • I'd like to do teratoma injections - saw from original hES paper that it was by injection into the mouse hindlimb muscle. Do teratomas form the same whether they are intraperitoneal, intramuscular, or subcutaneous? In the latter location, will we get a melanoma instead? Check with downstairs mouse people for protocol to follow to request approval for making human-mouse chimeras for research.

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.

  • Alethea 08:16, 23 June 2008 (UTC):
Personal tools