PDMS cell channel repeat
This is an update to my last post, where HUVECs mixed with collagen inside PDMS channels seemed to stay rounded up and not really do much of anything exciting.
This time around I made a few adjustments:
- New collagen protocol – gives 1.5 mg/mL solution before mixing with cells:
-10% 10x PBS
-1.4% NaOH
-38.6% DI water
-50% 3 mg/mL collagen - pH tested the collagen before cells and collagen after mixing with cells – pH was 7.5
- Aimed for higher seeding density
– paper claims to have used 6 x 10^7 cells/mL
– Confluent T25 typically has 3 x 10^6 cells total.
-actual seeding density = 5.5 x 10^5 cells in 0.75 mL
Results:
I also performed a live/dead stain 0n the 24-well after 24 hours to see roughly how many were still alive under these media-depleted conditions:
Things to try next time:
- Aim for more cells – grow flask very dense and suspend it in 0.5 mL
- try and up the final collagen concentration from 0.75 mg/mL to 1 mg/mL