I have two basic questions to ask about maintaining continuous cell lines.
1.) Is it harmful to a continuous cancer cell line if trypsinized 24 hours post splitting (if the split ratio while splitting turns out to be too high), even though the cells have attached well and show good morphology. The doubling time for the cells being 12 hours.
2.) How long should continuous cancer cell lines be maintained in culture post revival from liquid N2 before cryopreserving again without affecting its viability?
1. Yes it is harmful if you are trypsinizing your cells continuously after 24 hrs of splitting.It is advisable to do splitting after 48 hrs of splitting. I am also using lung cancer cell line. Initially continuous tryspinization may not affect the morphology. but after some passages you can see the difference in the morphology.As trypsinize is like a shock to cells and does not occur in natural conditions , it is advisable to do as less trypsinization as possible.For this you can seed less number of cells ( depends on your cells doubling time, for my cells it is 8 Lacs).
2. You can maintain cells until the morphology is good ( It depends on your cell type, some cells can go up to 80 passages and some up to 10 ). You can freeze them whenever they are healthy and with good morphology.
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