It's time to passage my iPS cells, so I add 1 mL Accutase per well for 5 min at room temperature (edge has folded back). But after aspirating Accutase and rinsing with fresh mTeSR1, I observed that undifferentiated cell colonies are still attaching.
In the below link, it mentioned that "differentiated cells are more strongly attached than undifferentiated cells", although I didn't find its reference source.
https://cellculturedish.com/release-your-cells-a-simple-new-tool-that-eliminates-the-need-for-manual-selection-and-scraping-in-hpsc-passaging/
My question are: (1) why differentiated cells release first after treated with Accutase? (2) whether I distinguish the cell differentiation correctly? Here, I regard fig 1 as differentiated cells, and fig 2 as undifferentiated cells.
Thanks for any answers or suggestions on this issue.
ReLeSR, discussed in the link, is non-enzymatic (they do not give a mechanism of action). Accutase is an enzyme that has proteolytic and collagenolytic activity. So, they have different specificity.
The link describes ReLeSR, not Accutase. If you want preferential
disassociation of undifferentiated colonies, then use ReLeSR.
thank you both very much for the answers Ron Hrstka and Ellen A G Chernoff . I have used ReLeSR to detach the undifferentiated iPS cells, it does work well.
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