Greetings, this is my first time working with Jurkat cells (https://www.atcc.org/Products/All/TIB-152.aspx#generalinformation) . I recently thawed out and cultured a vial of Jurkat cells in RPMI 1640 + 10% FBS. I believe I will passage them either today or tomorrow. I am following this protocol that I found here (copied below), however I noticed that nothing was mentioned of the use of trypsin or any other dissociation agent. My Jurkat cells have formed small, clumpy cell aggregates (as I see most do). Therefore in order to properly count them in single cell is trypsin required or is simply pipetting up and down enough?

Thanks!

https://www.researchgate.net/file.PostFileLoader.html?id=578731483d7f4bdcac712f78&assetKey=AS%3A383637720715265%401468477768430