I'm just started my training on glioblastoma cells. I want detailed about the principle and protocol for this particular search with explanation. To know deeply about it. Thankou
You may use any protocol you will find by googling "MTT assay". Eg. https://www.abcam.com/kits/mtt-assay-protocol or https://www.atcc.org/~/media/DA5285A1F52C414E864C966FD78C9A79.ashx
MTT assay is a very simple method, just find the right number of cell to be seeded in a plate before experiment. If you use U87 cells and your assay will take 72 hours, I would suggest to seed from 3000 to 5000 cells/well in a 96 well plate (but you should find it by yourself, as the cell growth may change due to many factors).
All you need to know about viability assays—including MTT—can be found in this chapter, available for free in pubmed bookshelf.
https://www.ncbi.nlm.nih.gov/books/NBK144065/
Thankyou all of you for your valuable answer. Now I know about MTT and its role in cell culture.
If possible can anybody help me in explaining my training topic.
Topic- Efficacy of Moringa olifera hexane extract on glioblastoma cells
Harish, M. oliefera has several therapetic properties such as antiancer, anti-inflammatory, anti-diabetic , antioxidant etc. Since you are interested in checking its effects on cancer cells (GBM), you can look into the molecular basis of its tumor suppressive activity. Additionally, most studies have been conducted using solvent extracts of MOL and not their soluble extracts. You can go for the latter one.
you treat your cells accordingly. At the end of your treatment, give it a PBS wash and then add MTT reagent at 0.5mg/ml (final concentration in respective media). Incubate the cells in dark in humidified 5% CO2, 37 degree incubator for 4-6 hours. Discard media carefully without disturbing the formazan crystals formed at the bottom. Add 100 ul DMSO to each well, dissolve the pellet completely and then take the Abs reading at 570 nm in any microtitre plate reader.
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