Event Title

Comparison of RAW 264.7 Cells to Murine Bone Marrow Derived Macrophages

Location

Allegheny A, Hotel Madison, JMU

Start Date

4-5-2019 5:10 PM

Description

Direct comparison of Bone Marrow Derived Macrophages (BMDM) to the murine derived RAW 264.7 cell line has not been done. Bone marrow from female Swiss mice is collected, and stem cells isolated for stimulation with M-CSF (PeproTech), to produce BMDM for direct comparison experiments. BMDM and RAW 264.7 cells are seeded at 4x10^5 cells per well in 500 uL of complete DMEM. After acclimation overnight, cells are stimulated with 0, 1, 10, and 100ng/mL of LPS for 24 hours. A sample of supernatant is used in the Greiss reaction to determine nitric oxide production as a measure of inflammatory function. Initial experiments comparing BMDM saw RAW 264.7 cells with a 10-fold greater level of nitrite production. Trypsinization of treated wells, after sample collection, is used to count cells in each well to normalize the response and better compare the two cell types.

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Apr 5th, 5:10 PM

Comparison of RAW 264.7 Cells to Murine Bone Marrow Derived Macrophages

Allegheny A, Hotel Madison, JMU

Direct comparison of Bone Marrow Derived Macrophages (BMDM) to the murine derived RAW 264.7 cell line has not been done. Bone marrow from female Swiss mice is collected, and stem cells isolated for stimulation with M-CSF (PeproTech), to produce BMDM for direct comparison experiments. BMDM and RAW 264.7 cells are seeded at 4x10^5 cells per well in 500 uL of complete DMEM. After acclimation overnight, cells are stimulated with 0, 1, 10, and 100ng/mL of LPS for 24 hours. A sample of supernatant is used in the Greiss reaction to determine nitric oxide production as a measure of inflammatory function. Initial experiments comparing BMDM saw RAW 264.7 cells with a 10-fold greater level of nitrite production. Trypsinization of treated wells, after sample collection, is used to count cells in each well to normalize the response and better compare the two cell types.