Dear all,
can anyone suggest a quantitative assay (not optical) to compare the viability and proliferation of cells encapsulated in different kind of hydrogel (alginate modified with various method)?
DNA quantification (PicoGreen) doesn't give consistent results since also dead cells are counted with the test. Evaluation of metabolic activity (AlamarBlue) is misleading since there is no balance of metabolite between the inner of the gel and the supernatant. *
Thank you so much for any answer I will get.
* http://www.ncbi.nlm.nih.gov/pubmed/15738673
Hi, I've just performed a similar test. Well, I dissolved my alginate beads with citrate buffer. Subsequently, I pelleted the cells and I resuspended the pellet in complete medium. Then, I performed a chemiluminescent assay for detecting cells proliferation.
Otherwise, I think you can dissolve the beads, pellet and suspend cells in medium, and after that, you can count cells with the Trypan Blue, in order to discriminate the viable from the dead ones.
Hi
You can try the Multitox-Glo Multiplex Cytotoxicity Assay. The kit is available from Promega. You can get quantitative information of both live cells and dead cells separately. The MultiTox-Glo Multiplex Cytotoxicity Assay gives ratiometric, inversely correlated measures of cell viability and cytotoxicity. The live cell protease activity is measured in terms of fluorescent emission, and the dead cell protease activity is measured in terms of luminiscent emission.
I am attaching the technical literature for this test. Hope you find it useful.
I always dissolve the beads, pellet and suspend cells in medium, and count the cells
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