Hi, I would like to measure proliferation in co-culture of MSC and leukemia cells, I have used CSFE but we would not like to detach the cells, somebody know how can I do it in a plate reader?
Thanks
there is specific plate reader called xCelligence that is the best device to measure proliferation with no staining or any other interventions , Please read here
https://aceabio.com/products/xcelligence-rtca/
xCELLigence Real-Time Cell Analysis (RTCA) systems is the real-time monitoring of cell proliferation, cell size/morphology, cell attachment quality, and cell invasion/migration. It is connected to specialized computer/software, so you do not have to make graphs manually using other programs, you get all graphs ready as you get the data.
But - you can use regular plate-reader (from BioRad or other). You need to stain cells with MTS ( so called tetrazolium reduction assay), see the protocol here
https://www.ncbi.nlm.nih.gov/books/NBK144065/
there are other assays available to cell growth analysis including use 5-bromo-2-deoxyuridine (bromodeoxyuridine or BrdU) which is a thymidine analog that incorporates into the DNA of dividing cells during the S-phase of the cell cycle.
Resazurin (Alamar Blue) is another example. Resazurin is a nontoxic dye that can also be used to measure migration and cellular invasion without resorting to sacrifice of the cells during the test procedure. Read this article:
https://www.ncbi.nlm.nih.gov/pubmed/22423572
I agree with the recommendation to use Resazurin/Alamar Blue/PrestoBlue for this. It's live cell-compatible, quick and sensitive enough to measure proliferation for most purposes. It's also cheap, which means you won't be too troubled to take measurements at whatever interval you wish.
Hi,
It's just a idea, and I haven't tried it by myself. Which cell would you like to know the proliferation in your co-culture? Both? If you can make sub-cell line from yours, probably you can make "secreted firefly luciferase" cells and "secreted renilla luciferase" cells. The secretion is for live-cell. I heard that Nano-luc reporter substrate can penetrate the cell membrane. It should be more clear.
Once you need to measure the luciferase activities of each single cell culture, then, you can estimate both cell proliferation from Dual luc assay of co-culture.
- Badges
- Science method