1. How should I store cryopreserved primary cells?
2. I've heard that properly thawing cryopreserved cells is an essential step in the culturing process. Could you give me some pointers?
3. How often should I change the media?
After thawing and plating the cryopreserved cells, the first medium change should be done after 24 hours or overnight, so that both residual DMSO and any dead cells are removed. Thereafter, the medium should be changed every 48 hours until the cells are ready to be passaged. More specific instructions are included in each cell line’s protocol.
4. How many passages can I obtain with primary cells?
For this purpose, the term "population doubling" is used instead of "passage" for describing the growth potential of the cells. A population doubling is a two-fold increase in the total number of cells in culture. A passage is the propagation of a cell population by subculturing from one vessel to 3 or 4 vessels. Because different researchers use different split ratios when they subculture, it is difficult to predict how many passages can be obtained with a particular primary cell. Our primary cells are guaranteed for a specified number of population doublings (as indicated within the product descriptions) when the corresponding cell-specific growth media are used for the culturing procedures.
5. How do I calculate the number of flasks or plates I should set up for culturing the cells?
The following equations can be used to determine the maximum number of culture vessels that can be set up:
The effective growth areas for common tissue culture ware are listed below:
6. Do I need to add any components into cell-specific growth media?
No, our complete growth media are fully supplemented and ready-to-use for your convenience. Each complete medium contains all the essential components for optimized cell growth.
7. Can I obtain basal media separate from the growth supplements?
Yes, we offer the basal media and growth supplements packaged individually.
8. What are the storage conditions and shelf life of the media and growth supplements?
For optimal cell growth please follow the handling instructions below.
9. Are there any special substrates or coating materials that I need to use for culturing Cell Applications' primary cells?
Most cells we offer can be plated on standard tissue culture grade ware. However, some cell types require coating material. For specific instructions, please see the respective cell product web pages to access the subculturing protocols.
10. When subculturing primary cells, what are the most important considerations to keep in mind?
Research. Development. Production.