Daniel:Notebook/HiResChrPaint/2013-9-10

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Fibroblast Adherence Timecourse

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Slides (From 09/09/2013)

9. Took photo at time point 21 hours

Results

File:Timecourse FinalComparison 090913.png

Even after 22 hours, the slides appeared to have few cells, and the cells present were not well attached. It is my theory that 1 of two things are happening: (1) The poly-L lysine slides are not providing strong support for the cells and they are not adhering; or (2) the cells are not getting enough nutrients. The second hypothesis is easier to test, so I am going to try that. If I can recover these cells with media, that will, I think, prove my point. Otherwise, I'll have to repeat the experiment. If I do that, I'd recommend:

  • Trypsin (1 mL)
  • Add DMEM to get to ~1 000 000 cells/mL, then add 100 uL to center of the slide
  • Wait 1-3 hours for basic adherence, then add 3 mL DMEM to promote more growth.


10. Added 3 mL DMEM to each culture dish (22 hours)

Media Addition Results

There was no improvement in the appearance of the slides at 3 hours post-media introduction, nor 6 hours post-media introduction. I didn't even bother to take pictures, the cells just looked dead. I think that they grew too long with only 100 uL media.