Oral Presentations
Location
Schewel 232
Access Type
Campus Access Only
Entry Number
6
Start Date
4-6-2022 10:30 AM
End Date
4-6-2022 10:45 AM
Department
Biomedical Science
Abstract
Parallel examination of the functional responses of bone marrow derived macrophages (BMDM) to the viral transformed murine RAW 264.7 cell line provides a basis for comparison of in vitro and ex vivo experiments using the cells as models for inflammation. Equivalent responses would allow the substitution of the RAW 264.7 cells for BMDM and reduce the use of animals in experimentation. RAW 264.7 cells are maintained in media and BMDM are generated by culturing bone marrow cells from female Swiss mice femurs. The cell lines are stimulated with lipopolysaccharide or E. coli, respectively, to induce an inflammatory response. Removal of the supernatant is used for quantification of nitrites in solution by the Griess assay. To correct for a slower growth rate of BMDM compared to RAW, cells will be dislodged from wells following treatment and counted by hemocytometer. Nitrite response levels will be normalized between the RAW 264.7 cells and the BMDM by establishing a response level per 10,000 cells for comparison purposes. Correcting for cell growth rates over the LPS exposure time will allow for better interpretation of results and comparison of experimental outcomes.
Faculty Mentor(s)
Dr. David Freier
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Functional Comparison of RAW 264.7 Cells and Murine Bone Marrow-Derived Macrophages (BMDM)
Schewel 232
Parallel examination of the functional responses of bone marrow derived macrophages (BMDM) to the viral transformed murine RAW 264.7 cell line provides a basis for comparison of in vitro and ex vivo experiments using the cells as models for inflammation. Equivalent responses would allow the substitution of the RAW 264.7 cells for BMDM and reduce the use of animals in experimentation. RAW 264.7 cells are maintained in media and BMDM are generated by culturing bone marrow cells from female Swiss mice femurs. The cell lines are stimulated with lipopolysaccharide or E. coli, respectively, to induce an inflammatory response. Removal of the supernatant is used for quantification of nitrites in solution by the Griess assay. To correct for a slower growth rate of BMDM compared to RAW, cells will be dislodged from wells following treatment and counted by hemocytometer. Nitrite response levels will be normalized between the RAW 264.7 cells and the BMDM by establishing a response level per 10,000 cells for comparison purposes. Correcting for cell growth rates over the LPS exposure time will allow for better interpretation of results and comparison of experimental outcomes.