Etchevers:Notebook/Genomics of hNCC/2009/07/03: Difference between revisions
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== | ==Finding the cells in vivo== | ||
As we only ever see what amounts to debris after 7 days post injection, we decided to try again using the currently cultured DRG cells, which are wildly dividing again in the 150 cm<sup>2</sup> flasks. | |||
Have stained 30 minutes with CMFDA 1:1000 after diluting two vials into 10.76 μL DMSO, warming to 37°C, then placing into 2x 10.76 mL warm PBS. Rinsed flask for EMB090005 DRG cells p8, replaced medium. | |||
Will aspirate, then replace with Rich medium for 30 minutes. Rinse with PBS, trypsin, spin down and count. | |||
Meanwhile, make up more dilute collagen: | |||
Diluted 78x (1.92 mL) into 150 mL 0.01M acetic acid (9 mL glacial) in MilliQ water, filtered into bottle. Then prepare a number of flasks for BWH group. | |||
*'''[[User:Etchevers|Heather]] 02:48, 3 July 2009 (EDT)''': | |||
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Revision as of 23:48, 2 July 2009
 Genomics of human neural crest cells
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Finding the cells in vivoAs we only ever see what amounts to debris after 7 days post injection, we decided to try again using the currently cultured DRG cells, which are wildly dividing again in the 150 cm2 flasks. Have stained 30 minutes with CMFDA 1:1000 after diluting two vials into 10.76 μL DMSO, warming to 37°C, then placing into 2x 10.76 mL warm PBS. Rinsed flask for EMB090005 DRG cells p8, replaced medium. Will aspirate, then replace with Rich medium for 30 minutes. Rinse with PBS, trypsin, spin down and count. Meanwhile, make up more dilute collagen: Diluted 78x (1.92 mL) into 150 mL 0.01M acetic acid (9 mL glacial) in MilliQ water, filtered into bottle. Then prepare a number of flasks for BWH group.
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