Any tips to dissociate fixed skin tissue?

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Jorge Morales Pedraza

According to kb.10xgenomics.com and thermofisher.com, dissociating fixed skin tissue into single cells can be delicate. Here are some tips that might help:

Use a suitable dissociation protocol: Tissue dissociation methods can impact the viability of the resulting cell suspension, so it’s important to pick a dissociation protocol suitable for your tissue type.

Unbiased tissue sampling: Bias in cell-type composition can be introduced during tissue sampling, impacting sequencing results.

Sample QC: Some sample metrics can be informative on the quality of the starting tissue and dissociation method.

Perform a pilot: If planning for a large experiment and handling a new sample type for the first time, running a pilot to test the experimental workflow and gain confidence before scaling up may be useful.

Optimal dissociation conditions: These should finely balance the cell release from difficult-to-digest tissue while avoiding cellular damage during mechanical and enzymatic digestion.

Mince tissue: Mince tissue into 3 to 4-mm pieces with a sterile scalpel or scissors.

Wash the tissue pieces: Wash the tissue pieces several times in a calcium and magnesium-free balanced salt solution.

Add dispase: Add dispase (0.6 to 2.4 U/ml in calcium and magnesium-free balanced salt solution).

Incubate: Incubate at 37°C for 20 min to several hours.

Remember, these are general tips and may need to be adjusted based on your specific needs and conditions.

Christine Pich-Bavastro

Thank you.

I am looking essentially for dissociation condition tips.

I have already improved different steps of the fixation/dissociation protocol (mincing tissue into 1-mm pieces, increasing digestion time, testing different enzymes, adding a mechanical dissociation step), and still doing in the pilots. But I was wondering if anyone has a good dissociation enzyme that dissociate small skin pieces and keep a good cell integrity.