I want to know after generating Knockout (KO) cell line of Human embryonic stem cell, how we can differentiate cell based on cell quality. Any specific protocol available which leads the specific results?
I’m not sure if you’re wanting to see like a morphological read out of a ‘sick’ cell, but if that‘a the case then maybe just stain for a membrane protein specific for your cell type(or something that’s know to be a part of this normal whatever cell) and compare KO and control. Or you Could use known markers for house keeping genes, or any proteins the field widely use to assess cell viability, proteins involved in the cell death or metabolic stress pathways
I am not sure, if I am getting your question correctly. Generally speaking, it is hard to morphologically differentiate between KO and normal hESCs unless your gene of interest has significant effects on hESC morphology, say causes a defect in cellular structure related proteins. Normally, clonal selection and characterization of multiple clones by PCR would be the best way to generate and develop a KO hESC line. Moreover, even if you where to morphologically identify KO vs controls, how would you isolate them or separate them for future work, as hESC grow as colonies?
Hope that helps.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line