Etchevers:Notebook/Genomics of hNCC/2009/03/24

Genomics of human neural crest cells

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Cell passages The hNCC from R1113 are doing very well and seem to have divided once in the dish, or at least spread to make whorls in the 35mm dishes. Counts on average for both 35mm dishes = 540K/2 cells = 270,000? seems few, given density. Plated into two collagen-coated 25 cm2 flask. Collagen - diluted 78x into 150 mL 0.01M acetic acid in MilliQ water, filtered into bottle. Keep in fridge. Placed 2 mL for 1h at 37°C on flasks. Took 12x 14mm coverslips out of 70% EtOH, placed in middle rows of 24-well Falcon plastic, and also coated in collagen. Prepared 4x flasks but only used two. Rinsed the others in PBS as well but left the 10 mL PBS on and stored in plastic bag in fridge. For the non-coated plastic, 400K/2 hNCC cells or 200,000 cells. Plated into non-coated 25 cm2 flask. Trypsined also all the neural differentiation cultures. Lots of cells recovered from the vitamin A + extra EGF cultures, clonal density for the FGF2/ extra EGF medium. Stopped trypsin in 10% FCS/DMEM but recultured in their non-serum-containing medium. Heather 08:06, 24 March 2009 (EDT):