Question: How do I prepare adherent cell lines for Single Cell analysis?
Answer: For adherent cells, enzymatic treatment is necessary to dissociate cells from the culture surface. This can be done using Trypsin, Collagenase, Dispase, TrypLE, Accumax, Accutase, or Trypsin-EDTA. The dissociation time varies depending on cell type. It is important not to over-incubate because this could damage cells. We recommend periodically checking the cells under a light microscope for complete detachment; the detached cells should appear rounded.
When using trypsin, the approximate dissociation times for some cell types are listed in the table below.
We have a Demonstrated Protocol for preparing single cell suspensions from cultured cell lines here: https://support.10xgenomics.com/single-cell-gene-expression/sample-prep/doc/demonstrated-protocol-single-cell-suspensions-from-cultured-cell-lines-for-single-cell-rna-sequencing
A more general Cell Preparation Guide that may also be helpful is available here:
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