Etchevers:Notebook/Genomics of hNCC/2009/03/02

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Cell consolidation

Re-fed the "N2" and "N2 + retinyl acetate" identical-appearing cultures with the last bit of medium (with 10x supplements). Had ordered B-27 supplement from the store, still not received, but I suspect it will be soon. The cells are looking good and fairly dense but not more confluent than 80-90% still.

Fed the Stemline+serum and new Rich medium 150cm2 flasks. Cells are looking good, not too much death, and rather smaller for the moment than they had been. Calling these passage 11.

As for the 4x 35 mm dishes from p8-9 and the two Rich and Stemline+serum flasks from "p10", there are a fair number of cells in all of them, more so in the Stemline+serum flask. Re-trypsinized all of them with a fresh bottle. Even after nearly 10 minutes, the latter did not unstick all the cells, but I noticed that they all moved as if in jelly. So I think the collagen coating was too thick (2h deposit) and the hNCC cells hunkered down in it.

I used a fire-polished pipette to "scrape" them off with jets of incident high pressure medium.

Consolidated:

  • the Stemline+serum and two 35 mm dishes (which had been re-fed with this medium) into one 15 mL tube. Cell count 115 x 104 / 2 cells (575,000 cells) added to the Stemline+serum 150cm2 flask with its original 550,000 cells for a total of 1,125,000 cells.
  • the Rich medium flask contents and the other two 35 mm dishes fed with Stemline+serum into another 15 mL tube. Cell count 86 x 104 / 2 cells (430,000 cells) added to the Rich 150cm2 flask with its original 1.02 million cells for a total of 1,450,000 cells.
  • Heather 05:40, 2 March 2009 (EST):
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