Etchevers:Notebook/Genomics of hNCC/2008/11/14
|Genomics of human neural crest cells||Main project page|
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After about 5' of trypsin with inefficient detachment, re-applied 0.5 more ml to the 35 mm dish and put in incubator for a few more minutes. Cells as retract leave a fair bit of space between them, which is why counts are not high - they occupy a large surface and seem to be growth-inhibited upon contact.
Stopped reaction in 7 ml of Rich medium (prepared now 2 months ago!) and spun down, then resuspended in 100 μL Rich medium. Placed 50 μL in 1 ml Rich medium in a new 35 mm dish, and diluted remaining 50 μL in Neural Stemline (Sigma, "N") medium to 450 μL. Counted this, and got for the entire surface (9 fields) of counter 69 cells, and these in some clumps of 2-5 cells.
This means that there are some 76500 cells total in the original dish (or perhaps lost some) and 38250 cells per dish now. Or thereabouts. See if they survive the weekend.
Submitted my two evaluations for the Georgian National Science Foundation.