Hello,
Today I want to ask several questions about cell works.
I am just a junior in the field of my research, so compared to others,
I may ask repetitive and too monotonous questions but I want all of you
to understand me.
I always realize it is impossible to handle even all cells from one specific culture tool(It can be dish or well plate or other tools) in the totally same way during all cell works, including cell splitting.
So
I wonder whether it is ok to simply consider all cells in same culture tool are considered to be in
same specific cell passage number and in same condition.
Thank you!
If you "seed" cells at the same concentrations (cells/ml) in different plastics you will find same conditions. Generally the difference are: 1) the volume of the media you have in your petri or flask, 2) the medium of trypsin or PBS you use to detach cells.
Plastics with different sizes will produce different amount of cell if you use the same inoculum concentration due to the different size. For example if you plate cells at 10.000 cells/ml in a T25 flask you can use 5 to 8 ml of medium while if you use a T75 flask you will use 10 to 15 ml. it is obvious that the number of cells you added is different and so after 1 or 2 days you will have as absolute value more cells in T75 than T25.
Dear Diego Baderna,
Thank you for your reply!
I am impressed with your knowledge.
Yes, size of culture tool is very important. I absolutely agree with that.
So I wonder whether newly seeded cells from one culture tool or culture tools of same passage can be considered to be in same condition and in same passage if they are cultured in same-sized tools, although I can't handle all these cells totally in the same manner during split and next works.
If you don't mind, can you answer for my additional question?
Thank you agian!
Hi, I hope I understand your doubts in the last reply: "I wonder whether newly seeded cells from one culture tool or culture tools of same passage can be considered to be in same condition and in same passage if they are cultured in same-sized tools"
If you seeded your cells for example after thawed them from a cryovials, or you seeded your cells (same pasages) after detached with trypsin in two or more petri (or flask), yes they can consider the cells in the same conditions.You have to pay attention how to handle cells because thay have to be in same good conditions in particular when you have to repeat experiments during several weeks.
Commonly, when you work with cells, you have two T75 flask (someone prefer the 10mm petri or 150mm the big one), so you have a lot of cells when you need them. You count the cells for your experiment and seed again the rest in the big flask or petri, in general during the first passage if remain cells after the experiment and the seeding it is better to freeze a good number of vials.
- Badges
- Science method