I am currently working with HepG2, HepAD38, Huh7 and HepG2-NTCP-K7 cell lines. I want to know what are the correct timeline for those cell lines.
Everything depends upon the initial seeding density. You must also refer to the seeding density from the source from where the cell line is procured.
Hello Kamindu Gayashan
I have not worked with these cell lines. You need to find an answer yourself by either doing a literature search for these cell lines or performing some pilot experiments. I could suggest a few points that could help you.
Every cell line has a doubling time. Check for the doubling time for each of these cell lines. However, the doubling time may differ depending on the culture conditions you subject the cells to.
Cells should be passaged when they are 80% confluent. Ideally, cells should be split at this stage because this will improve overall cell viability. Splitting your cells at this stage will also lead to less aggregated cells. These cells will also have a shorter lag time (i.e., the time taken before cells start logarithmic growth) after they are split.
If you passage cells at high confluency, it may dramatically affect cell behavior and culture kinetics. Cell death can happen at high confluency because nutrients in the media become depleted or cells start competing for space on the culture dish or flask surface.
Another point which you need to consider while culturing these cells is the seeding density. As Deepti Mittal rightly mentioned, seeding density plays an important role. Seeding the right cell density is necessary for optimum cell growth. Seeding a lower cell density will result in lack of cell to cell interaction and communication. As a result the cells will show slow growth and eventually die.
On the other hand, seeding a high cell density than the optimum will result in cellular stress as there will be overcrowding and lack of space for cells to interact with each other and grow. Moreover, nutrients in the culture medium will be insufficient and there will be accumulation of metabolic waste causing cells to face stressful conditions.
You may take these points into consideration when you culture these cells.
Good Luck!
These cells vary in populations doubling time — some divide faster than the other which is related to their biology and media composition. So I suggest do a population doubling time calculations based in the initial seeding cell density and yield cell density (until 70% confluency). But make sure to change the media every two days.
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