Hi all,
I ran into a problem where if I smash the thymus on a petri dish first, then filtering them through a cell strainer, I would get better quality of cell pellets. However, when I smash my thymus directly in the cell strainer, I would get weird cell pellets: some of the cells form a thin layer on the side of the 50 mL Fecon tubes. Could I be hurting the cells somehow when smashing the thymus directly in the cell strainer?
Hello Yinghua Wang
You could follow the below protocol.
Put the thymus into a petri dish and add 10 ml PBS to the petri dish. Place a cell strainer in the petri dish with the thymus (do not place the thymus in the strainer until it has been sliced). In the petri dish, hold the thymus with the tweezers and slice the thymus with the scalpel. Repeatedly cut the thymus until it is in 1 cm chunks and place the chunks into the cell strainer in the petri dish. Hold the cell strainer and mash the chunks to filter the cells into the petri dish and create a cell suspension in PBS. The PBS in the petri dish will turn cloudy as cells are released into it. Then pipette the cell suspension into the 50 mL centrifuge tube through the cell strainer to further remove any clumps.
Best.
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