Hello!
I know cell viability is the main concern when animal cell cultures are stored at -80°C for long, but what other negative effects do you know of? Any information at the cellular as well as genetic/protein levels is greatly appreciated.
Kind regards,
Shahin
Mainly viability and all of the issues that come about when you gradually lose viability. Some other issues: 1) Proteins and enzymes (proteases, DNAases, etc) are not as inhibited in -80 which means they will continue to damage the cell, both DNA and proteins. 2) Subclonal populations will take over. So specific subpopulations that do not represent the original cell lines which are more stable at -80 will live longer and grow out, 3) Cytotoxicity will occur from all the cellular lysis, causing massive cell death upon thawing. 4) Inconsistent data sets. It is very unlikely that 2 vials of parent stock will thaw out with the same cell populations, as clonal survival will be quite random 4) Loss of heterogeneity, genomic and proteomic. Mammalian cell cultures is a group of cell populations, not one. As cells die off you will start loosing that heterogeneity and be left with clones that exhibit strange and unreproducible genomic and proteomic characteristics.
Mainly viability and all of the issues that come about when you gradually lose viability. Some other issues: 1) Proteins and enzymes (proteases, DNAases, etc) are not as inhibited in -80 which means they will continue to damage the cell, both DNA and proteins. 2) Subclonal populations will take over. So specific subpopulations that do not represent the original cell lines which are more stable at -80 will live longer and grow out, 3) Cytotoxicity will occur from all the cellular lysis, causing massive cell death upon thawing. 4) Inconsistent data sets. It is very unlikely that 2 vials of parent stock will thaw out with the same cell populations, as clonal survival will be quite random 4) Loss of heterogeneity, genomic and proteomic. Mammalian cell cultures is a group of cell populations, not one. As cells die off you will start loosing that heterogeneity and be left with clones that exhibit strange and unreproducible genomic and proteomic characteristics.
Daniel, thank you very much for your thorough response. Have a wonderful day!
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