I started with a known amount of cells suspended in matrix beads but once the cells have grown into spheroids, they are of different shapes and sizes. In order to normalize the response between different experiments I need to be able to quantify cell number and viability. I would appreciate suggestions that are practical for routine work. Thanks.
Hi Barae,
There is a product from Promega designed for measuring cell viability in 3D cultures that it may work for you: CellTiterGlo 3D Viability Assay.
Another option that I can think about is the RealTime-Glo™ Assay (Promega), that allows you to measure viability over 72 hrs. I wonder if you could add it with your cells on Day 0, and keep measuring the viability after that..
Lastly, you could potentially correlate cells# over time doing DNA quantification. I would extract the DNA just the way you would do with a tissue sample, and compare the amount of DNA among the time points.
I don't see how to separate the cells to count them.. Anyone?
Best,
Marilia
Hi Barae,
using microscopical technique this kit from SIGMA is great to evaluate live and dead cells: Live/Dead Cell Double Staining Kit (Calcein AM and Porpidium).
Good luck
Hi Katerina,
Thanks for the lead. That sounds like very interesting research that you did but I couldn't get the whole text, could you send me the full text. How did they calculate the growth rate of the entrapped cells?
Indeed there are two things that have to be considered. First I will have to perform a cell viability assay like the Live/Dead Cell Double Staining Kit mentioned by Cesare. Then I will have to follow up with dissolving the beads with sodium citrate and count them under the microscope or with a cell counting device such as the Coulter counter. Thanks for the help, the file and the links!