Hello everyone, I need to perform a culture of lymphocytes isolated from healthy volunteers' blood samples using Ficoll Paque and that in order to study the DNA damage induced by several chemotherapy agents as well as DNA repair capacity. Therefore, I need to know if there is any optimal protocol that allows good PBL survival ? Also, for how long can we keep the lymphocytes in culture before doing the drugs tests ? Thank you
Hi Iman,
The big problem is that PBL consist of many cell types. Each cell type has its own optimal culture conditions. (T-cells have other requirements than NK-cells). This is especially the case if you want to expand your cells.
Culturing cells often has an influence on the expression of proteins. This might influence the results of your experiment. So my advise would be to culture the cells as shortly as possible.
With kind regards,
Ronald
We used medium199 supplemented with fetal bovine serum and harvested between 69 and 72 hrs.It was good for whole blood and chromosome analysis
Hi,
Please see the following link
Article Current Protocols in Cell Biology
Dear
You can follow this article:
Lefort CT, Kim M. Human T lymphocyte isolation, culture and analysis of migration in vitro. Journal of visualized experiments: JoVE. 2010(40)
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