I plan to do the CRISPR-cas9 base edit in mouse cells, it is better to use the primary cells or immortalized cell lines? I have read some papers, some used the primary cell lines.
Hello Shasha Alice Cheng
The cells used in CRISPR-Cas9 should be stable, predictable and easy to handle. The immortalized cell lines have been developed to survive in culture, which can cause them to differentiate from the human tissues they are supposed to represent. Immortalized cell lines have acquired the non-physiological ability to proliferate indefinitely due to accumulated or induced genetic mutations. Continuous passage of immortalized cell lines over time may affect their original physiological properties, resulting in misleading results in studies.
On the other hand, primary cells are more physiologically relevant than their immortalized cell line counterparts. Primary cells, unlike immortalized cells maintain their biological identity and can only be propagated for a few generations in vitro. For screening purpose, it becomes difficult to use primary cells because it is hard to keep them alive long enough for the editing and invitro analysis as well as for the screening techniques.
Using alternatives that better resemble the natural state of the organism would be preferred. Therefore, it becomes important to focus on primary cell lines because primary cells will bring us one step closer to the model organism that are relatively easy to manipulate and assay, compared to the organism itself, making them the gold standard for studying human diseases.
Best.
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