My suggestion is to disaggregate the tissue with a cell strainer and a syringe plunger under appropiate conditions -which may vary depending on your tissue and what you would like to do with it afterwards. Once disaggregated you can store them frozen in presence of DMSO. It is advisable to measure the viability of the cells before freezing and after thawing with any method. I link here my method of choice.https://www.researchgate.net/publication/261139124_A_fast_low_cost_and_highly_efficient_fluorescent_DNA_labeling_method_using_methyl_green

Best,

D

Hello Kanchan.

Chop the tissue into small pieces 2~3 mm in diameter in collagenase (you have to select what type of collagenase depending on the origin of tissues) in Hank's BSS solution in 100 mm dish. You may need several number of dishes to avoid clogging and aggregation. Incubate the dish 3~4h in 5% CO2 in incubator. While you are collecting slurry without tissue lumps of BSS solution into 50 ml tubes. Spin them and save cell pellet, followed by Trypsin-EDTA sol (T-E) for 10 min while watching under mike. Also T-E the tissue lumps in an Erlenmeyer flak with a small gentle stir bar and usual cell collection procedure. Transfer the suspension into 20% FBS+Hank's to stop further digestion and usual cell collection procedure. Pour more T-E into the flask to make more cell suspension and repeat this procedure.

You should have single cell suspension to avoid cell damage during freezing and thawing, so care must be taken to observe single cell suspension, and not too long T-E digestion and quick stopping digestion transferring into FBS-containing Hank's.

Follow usual cell freezing procedure by adding gradual addition of DMSO and FBS. Even thawing procedure is critical for cell survival of primary cells when you need any analysis.

Regards.

Thank you very much Sang Ho Lee and Daniel Prieto for your answers. These will be really helpful for me.

Additionally, if you receive the tissue and can't process it right at the moment, you can save it in a reagent to preserve the integrity of RNA. In case you don't want lose the tissue, you can still make further expression analysis with it if interesting to your project.