I read several protocols about CD8 T cell culture, including using 1640 or aMEM with FBS or commercially established expasion medium, with IL-2 or IL-7 added and OKT3 activated. Their results show 100-1000 fold T cell expansion with high viability. But, following their protocol carefully, I only can expand my cells in a small number, with lots of cell fragments in the mudium. I can't find out why I cant repeat their results.
Dear Jaba Tkemaladze
, thanks for your answer. but I am confused if the temperature is a relative factor? I just placed the cells in the 37C incubator and never cared about the temperature. ps the cells were counted once every 2 days. they started at 2x10^5 and reached 2x10^6 on day 7.
Hi!
When do you remove cells from the stimulatory antibodies? At which step?
I worked on murine CD8 T-cells only. And the protocol was to stimulate them overnight with aCD3/aCD28, and then culture on IL2 or IL15 changing the culture medium or diluting cells once in couple days. Or even on IL15 only.
As for the human CD8 T-cells, I guess there may be a trouble with increased amount of effector cells, which may be more prone to AICD.
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