Poster or Presentation Title

Comparison of RAW 264.7 Cells to Murine Bone Marrow Derived Macrophages

Student Author Information

Delia LoSapioFollow

Access Type

Campus Access Only

Presentation Type

Oral Presentation

Start Date

April 2019

Department

Biology

Abstract

The convenience of the RAW 264.7 murine macrophage cell line makes it a common alternative for Bone Marrow Derived Macrophages in experiments investigating inflammatory functions of innate immune responses. However, direct comparison of 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 4x105cells 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 and RAW 264.7 cells 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.

Faculty Mentor(s)

Dr. David Freier

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Apr 10th, 11:45 AM

Comparison of RAW 264.7 Cells to Murine Bone Marrow Derived Macrophages

The convenience of the RAW 264.7 murine macrophage cell line makes it a common alternative for Bone Marrow Derived Macrophages in experiments investigating inflammatory functions of innate immune responses. However, direct comparison of 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 4x105cells 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 and RAW 264.7 cells 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.