I am currently working with human lung epithelial and fibroblast. I would like to know which passage is best for invitro studies as well. Kindly advise. Thank you
Hello Pavithra,
Primary cells are superior to established cell lines as they can act as model of the in vivo situation. But the disadvantage of primary cells is that they have limited life span and grow slow in culture. Moreover, with every passage in culture, the primary cells will keep changing its characteristics.
So, if you are going to use primary cells for invitro studies, I would recommend you freeze these cells at early passage after you find them growing well in culture. It is advisable to use primary cells below passage 10 for invitro studies. If you wish to use established cell line for invitro studies, try to use cells below passage 20-25.
Primary cells have a finite lifespan and will undergo senescence after a few passages in culture, while immortalized cell lines are cells that have been modified in a manner that they can be cultured indefinitely. Cell immortalization can be achieved by introducing viral gene to override the cell cycle or by spontaneous mutation or by any other similar technique.
Best.
Malcolm Nobre Thank you so much. Do u mind to share any supporting paper on this, if possible.
Generally with primary cultures the lower the passage number the better, as there is a reduced chance of accumulated mutations or other epigenetic changes.
I personally aim to use my primary lung epithelial cells at passage 3-4
Cells are generally considered primary cells until they have undergone a certain number of passages, which varies depending on the cell type and the laboratory. There is no strict rule for determining when cells become non-primary, but here are some general guidelines:
- Human diploid fibroblasts: These cells are typically considered primary until they have undergone around 10-15 passages. After this point, they start to lose their normal cellular functions and begin to exhibit altered growth properties.
- Human bronchial epithelial cells: These cells are typically considered primary until they have undergone around 5-10 passages. However, some labs may consider them primary up to passage 15 or higher.
It's worth noting that the number of passages alone does not necessarily determine whether cells are primary or not. Other factors, such as the quality of the culture conditions, the purity of the cells, and the presence of contaminating cells, can also impact the classification of cells as primary or non-primary.
For in vitro studies, it's generally recommended to use low passage numbers (e.g.,
Sam Siljee and Saif Wahid Thank you for the suggestion. Can you share papers regarding this, if there is any.
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