Etchevers:Notebook/STRA6 in eye development/2010/03/20

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Cell culture for CCE-Rx and CCE-EGFP

Second split of the cells. First round after 24h post-thaw - there were 20 x 10^6 in the CCE-EGFP 100 mm dish, and 10 x 10^6 in the CCE-Rax dish but the cell clusters were bigger. Maybe 90% confluence in places.

Today used TrypLE 5' and having seeded 50K cells two days ago, today there were 2,400,000 to 2,500,000 cells. So in two days, there was 50x expansion. Not bad!

Had frozen the rest of the others. 2x 10 million cells for CCE-EGFP in 50% serum, 40% medium, 10% DMSO. Perhaps too many but remember when you thaw. 2x 5 million cells for

Seeded 21 uL each into new dishes.

Collagen is ready to prepare flasks, at 100 mg for 3.54 mg/mL. Need to calculate the right concentration for coating Monday morning.

  • Heather 10:18, 21 March 2010 (EDT):