Does anyone have experience growing single cell clones of MCF-10A.I have generated gene knockouts and need to pick single cell clones from these cell lines. It is challenging as these cells are immortalized but pre cancerous and require a lot of supplements in the media to grow. One idea is to try condition media. Other ideas will be highly appreciated. Thank you!
Dear Abhay!
Please You look at the articles:
Article A novel MCF-10A line allowing conditional oncogene expressio...
Article Characterization of MCF-12A cell phenotype, response to estr...
Actually mcf10a cells dont need many other supplements to grow. I think additional sodium pyruvate can do the trick upon dmemf12 and fbs 10%. For selection of single clones there are many methods available. The most efficient in my opinion is the moniclonal selection by adding 1/2 cell per well in 96 well plate and wait for the cells to grow from a single cell. You might need conditioned media to achieve clonal selection as a single cell might not be able to grow in normal media and may require additional signals fromthe conditioned media.
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