Good day all, I am currently trying to familiarize myself with the culturing and maintenance of HL60 & JURKAT leukemia suspension cultures. What density should these cells be cultured at, how often should I split them, show should I split them? Should I be doing regular viability testing (eg. trypan blue staining)? Any guidelines or even publications regarding this would be appreciated.
Please go through following links:
http://www.atcc.org/products/all/CCL-240.aspx#culturemethod
http://www.atcc.org/products/all/TIB-152.aspx#culturemethod
Dear Alexis,
I have cultured these cell lines before. Jurkat grows in clumps so I did repeated pipetting to break the clumps before cell counting. I split the cells at 0.5x10^6 cells/mL every 2-3 days. You don't have to do regular viability testing but trypan blue is used anyway each time you split and count the cells.
Hi Alexis,
You should maintain cultures at 1-5 x 10^5 cells/ml. It is recommended that not culture them more than 6 week.(they will be differentiated). Also you can doing regular viability testing by 0.05% Trypan blue dye. You can wash the cells via 600-700 rpm/10 minutes by centrifugation.
Be succeed
yours
Amir
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