In
the last five years a series of protocols
has been developed to generate dendritic
cells in high quantities. In our country,
research in this area of the immunology
has been initiated with the intention of
using them in therapeutic approaches, in
particular in developing experimental protocols
for immune therapy against cancer. These
studies require to be validated and deeply
studied using experimental models, particularly,
the murine model. In this project, the primary
goal was generate murine dendritic cells
derived from stem cells from the bone marrow,
using for this issue an experimental protocol
standardized by Lutz and col. (1999) and
modified in this study. Then, for the phenotypic
characterization of dendritic cells, surface
markers such as; CD11c, CD86 and H2Kb were
analyzed, through quantification by flow
cytometry. The final goal of this work was
to standardize a method for obtaining murine
dendritic cells with the stable phenotypic
characteristics, to be use in the immunization
of mice with antigenic peptides for the
study of biological mechanisms in the tumor
immunology. |
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The results obtained at different days of
cell culture (6, 8 and 10 days), showed
a high presence of molecule integrin CD
11c and the coestimulatory molecule of T
lymphocytes, CD86, at days 10 of culture,
which would indicate a estate of phenotypic
maturity of analyzed dendritic cells. In
addition, it observed a high expression
of Mayor Histocompatibility complex class
I molecules (H2Kb), in analyzed dendritic
cells, indicating that those cells may be
able to present antigen and induce a specific
T lymphocytes response in vivo. Finally,
a double cellular staining was performed
to detect the population that co-expressed
the molecules CD11c and CD86 and which we
defined as dendritic cells. The result showed
that 55.27% of produced cells should correspond
to mature dendritic cells. These studies
will allow the initiation of project about
T lymphocytes in vivo induction through
the immunization with in vitro established
dendritic cells.
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