Dear all,
I genetically engineered the human primary cells with CRIPSR-Cas9 system and currently can passage them stably in vitro. The NGS analysis showed multiple mutations of target sites but not wild type suggesting the presence of multiple clones. Can we call them a cell line? or, only monoclonal cells should be called a cell line?
Thank you in advance!
Do a single cell clone and expand..!! It might take a month time or little more. Expand and freeze multiple vials as needed and do a confirmation at the time of freezing vials to make sure it retains the change but don't have other mutations observed..
Hi Goodwin,
Thank you for your suggestion. We need to analyze multiple aspect of the engineered cells such as RNA expression, karyotype, or other things, but I thought characterization using the multiclonal cells would only show average data of each clone, I was hesitating to do so.
Thanks!
Yes, multiclone cells change their phenotype+genotype over time because each time when you split the cells, there is a chance for loosing cells with certain phenotypes/genotypes.
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