I had cultured primary human lung cancer cells derived from needle biopsy sample after isolation. For 10 days cells were greatly healthy and good in number. I had two different cells from different patient sources. Both got detached from their respective flask & petri plate at the same time within 24 hours after 10 days of successful culture. I am unable to understand the cause behind it.
Hello Pragati Sharma
It is important to retain the viability of primary cells after isolation as most of them undergo the process of senescence and stop dividing after a certain number of population doublings. Therefore, for long-term viability of the cells excellent cell-culture handling skills along with appropriate culture condition (i.e. growth medium, temperature, gas mixture, pH, presence of proper nutrients) are essential. You need to include antibiotic in the growth medium to inhibit contamination introduced from the host tissue. However, long-term use of antibiotics is not recommended. Insufficient or absence of attachment factors in the growth medium or contamination or over exposing cells to trypsin during trypsinization could probably be the cause of the problem.
Also, the characteristics of primary cells changes with each subsequent passage if optimum culture conditions are not maintained.
Best.
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