Should I use warm or cold tissue dissociation?

Question: Should I use warm or cold tissue dissociation?

Answer: Numerous tissue dissociation methods are available, each with its advantages and disadvantages. For fresh tissue, many researchers use enzymatic dissociation, which can provide high cell yield and viability. Many different enzymes are available for dissociation, and resources exist to match the best enzyme to your tissue and organism of choice. Keep in mind that some enzyme treatments may modify cell surface proteins and therefore would not be compatible with Feature Barcode technology for Cell Surface Protein, flow cytometry, or other orthogonal assays. Enzymatic dissociation can also be time-consuming. An alternative is a mechanical dissociation, which reduces processing time and works well with easy-to-dissociate sample types. However, cell recovery efficiency and cell viability can vary across users and experiments. Mechanical and enzymatic dissociation can be coupled together.

Recently, researchers have identified differences in cell-type composition and gene expression between enzymatic and mechanical dissociation for certain sample types. O’Flanagan et al. (2019) found that using enzymatic dissociation at cold temperatures helped minimize stress responses during mouse tumor dissociation. Mattei et al. (2020) observed differences in transcriptional profiles of cells isolated from brain tissue using enzymatic dissociation at 37°C or mechanical dissociation at 4°C. 

When deciding whether enzymatic or mechanical dissociation is best suited for your experimental goals, consider existing knowledge of how each of these dissociation methods may impact your cell type of interest. If there is no specific dissociation method for your cell type available, consider assessing differences in gene expression, cell surface protein expression, and cell-type distribution by performing a side-by-side comparison of each method in a low-throughput experiment using Chromium Single Cell Gene Expression LT. An informed decision can then be made based on empirical data before scaling up to additional samples.

A few known examples of cold and warm dissociation on different sample types are shown below:

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