I treated the cells , BY2 cells, with an organic material but they grow like the control while, I expect the better grow and, this problem is driving me crazy as I'm doing it for about three months! I though that probably apart from the concentration of the treatment, there must be sth else prevent the cells to respond appropriately; I don't know sth like the number of the cell under treatment. So, if it's the case, I need to make sure that the cells are growing, suspension culture, perfectly and ready to go to the next step like applying treatment. I mean, I need to have a clean baseline for the growth. Since depending on the initial cell number, the final concentration is different. I want to have the same amount of cells after every sub-culturing. However, I am not sure if it makes any difference or not. However, counting is such an untidy work as the plant cells are stuck together.
any idea?
best,
Dear Babgohari,
I accept that it is not an easy task to count the number of cell for suspension culture like BY2. You can check the effect of your supplement on the growth of your BY2 culture by calculating “Packed cell volume” of culture. Increase in PCV is proportional to the increase in number of cell in the suspension. It can be easily calculated by spinning the BY2 suspension culture in 50ml falcon tube. After centrifuge, the height of the BY2 cell column (pellet) is measured and compared to the total height of the column of media (supernatant). The percentage of the total volume occupied by the BY2 cell mass is the Pack cell volume (PCV). (PCV) depends mostly on the number of BY2 cells. But there is some effect (to a much lower extent) of the average size of the suspension cell in different experimental setup, which can be checked using microscopic software. The use of swinging bucket rotor is recommended for PCV calculation.
I hope this may help you..
In research, sometimes, when you see a specific cell line (with a specific genotype) somewhere which reacts to a specific compound, it does not mean that different cell lines (in this case, BY-2) will have the same response to the compound. They may not response to it at all or with minimum response. Hopefully this is not your case, but you need to take this possibility into account, and have a backup plan. Have this compound been tested on BY-2 before?
Thanks for you answers,Yeah dear Yuan-yeu you're right it's possible that it doesn't make any difference, and probably i need to focus on other aspects and cell analyses to see the changes, not just the growth. That compound has not been used for BY-2 cells.
best
- Badges
- Science topic
- Similar topics
- Indexes