Etchevers:Notebook/Genomics of hNCC/2009/02/27

Genomics of human neural crest cells

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Cell exploits First, have just ordered B27 supplement (Invitrogen) because it's in the store, and because I only have enough N1+Stemline Neural +/- retinyl ester supplement as per last week, to last for another couple medium changes or so. And the one with retinyl ester is pretty close to confluent - not looking neuron-like - so may need to be passaged. Second, have trypsined the Rich or Stemline Neural + serum 25cm2 flasks and the first trypsined much better than the second. Inefficient rinsing? There didn't seem to be an initial density difference, but there were 2x as many cells in the first as in the second: Rich = 204 x 104 cells * 0.5 mL = 1.02 million cells, seeded in 150 cm2 flask. A bit low density, perhaps. Stemline neural + serum = 110 x 104 cells * 0.5 mL = 550,000 cells. Definitely low density. But this culture is less important. It would imply that not all cells doubled since last week, whereas in the Rich medium, they would have done. Or some of them divided three times and others not at all... Heather 09:30, 27 February 2009 (EST): I am such a noodle, as they say in French. Here I was wondering how I ran out of N-1 supplement so fast. I only diluted it 10x instead of 100x! Do better with the B-27 supplement, okay? (use up the N-1 first though, as I can not just dilute 10x [| because of the other adjuvants]). Made fresh Rich medium using a frozen aliquot of the new ES serum. Heather 10:04, 27 February 2009 (EST):