I am currently looking to purchase some viability assays for my research which involves testing the viability/attachment/toxicity of breast cell lines such as the MCF-12A to different manufacturing scaffolds.
I have read that most papers use kits such as MTT, calcein and ethidium and resazurin. However, most of they do not specificate the specific kits.
Another concern of mine is the solubility of these reagents in gel like scaffolds? For example MTT use in matrigel, which I have read that can be challenging.
Are these the best methods? And if so, which kits would you suggest?
Thank you for your time!
I would suggest FACS where you have to perform each testing like estimation of cell viability, estimation of drug resistance and toxicity , analysis of apoptosis.
Other than this Trypan Blue assay is also helpful for cell viability.
Dear Luana,
working with cells in scaffold matrices and determining their viability status can be quite challenging. Therefore - for a first (and quite cheap) analysis - I would suggest to simply go for life/dead analysis using the fluorescent dyes DAPI (or Hoechst 33342) and propidium iodide. Diffusion in scaffold materials is no problem (at least in my experiments) and washing to remove unbound dyes can be kept at a minimum since fluorescence can only be detected when dyes are bound to double stranded nucleic acids in the cells.
Sometimes DNA molecules present in scaffold matrices could cause some headache, but that should be only a minor issue.
All the best with your experiments,
Christian
- Badges
- Science topic
- Similar topics
- Physical Sciences
- Materials Science
- Materials
- Composites
- Nanocomposites