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
Dear Alexis,
I have cultured U937 cell line before. A good news is that, they will be visualized by phase contrast microscopy and the number of apoptotic and necrotic cells could be estimated according the appearance of the cell and you will not be in an a force to follow the time consuming trypan blue staining processe.
Good Luch in your research
Hoda
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