Blood cells
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As stated before blood is probably the most accessible source of cells to do research on. The number of cells is large and it is not too inconvenient for the donor. A lot of research questions is build around a peripheral blood cell model. The larger populations are the lineage populations like the lymphocytes (B and T), the monocytes, NK cells and the granulocytes (basophil, eosinophil and neutrophil). And than there are several other cells like the uncommitted CD34+ cells and blood dendritic cells. But these are found in very low percentages.Depending on the question peripheral blood mononuclear cells (PBMC) are isolated via density centrifugation. This way all red blood cells AND the granulocytes are lost in one isolation. Looking at a more physiological situation it is possible to use a lysing solution (see protocols) before or after stimulation of whole blood. That way only the red blood cells will lyse and all other (white) blood cells will be left. It is also possible to look at the in vivo situation by staining the cells directly. Expression of cytokines in cells found in bronchial alveolar lavage fluid, the levels of granzyme and perforin in CD8+ T lymphocytes, cytokines in monocytes in sepsis or stimulate/manipulate the cells at forehand and look at antigen specific induced cytokine production by CD4+ T lymphocytes or lipopolysaccharide (LPS) induced cytokine production by monocytes. We will show some results concerning the T lymphocytes and the monocytes regarding cytokine staining. |
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