In order to check cell activity on hydrogel scaffold, which in vitro assay is recommend? Will the absorption property of hydrogel affect the mention assay?
Thanks.
MTS worked really well for us in hydrogel. It is the water soluble version of the MTT. You do not need to solubilize the dye with DMSO at the end of your assay.
See materials and methods here: http://www.pnas.org/content/110/32/12887.abstract
Hope that helps.
Hi,
Usually, what I do, if the cells are seeded on top of the hydrogel it would be easy.
Just make sure that you have the hydrogel alone as the background signal. I have used Alamar Blue and MTS and both were fine.
Measure the signal intensity for cells on the gel and then subtract the gel alone from the measured value.
Hope it works.
Alamar Blue is excellent - also means you do not have to destroy cell samples as you can measure the same population of cells at different timepoints; thus it is a more kinetic method of real-time growth/metabolism within the same cell population within a hydrogel.
Good luck.
Both MTT and alamar blue are very good compounds to use. Beware that for the end point MTT assay, you must remove as much of the cells from your hydrogels (if you are performing a growth study on the gels themselves), otherwise your viability count vs your controls will be very skewed.
Thanks everyone for the answer. Recently, I have problem with MTT like Kenny mentioned. I will try Alamar blue or Presto blue instead.
Hands down to MTT, I have tried and tested Alamar blue, however Both have pros and cons on there own. I would still advice and use MTT
Hi, I'm also experiencing problems using Alamar Blue on cell-seeded hydrogels in which the resazurin is not being reduced in hydrogel wells (no color change). Does anyone know a way around this?
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