Comparison of Two Standard Growth Media for the Inflammatory Response of Raw 264.7 Murine Macrophages
Location
Turner Gymnasium
Access Type
Campus Access Only
Entry Number
49
Start Date
4-5-2023 12:00 PM
End Date
4-5-2023 1:30 PM
College
Lynchburg College of Arts and Sciences
Department
Biology
Abstract
The RAW 264.7 murine macrophage is a significant cell line for the study of inflammation and one of the most extensively used cell lines in mammalian cell culture. The standard medium for its growth and maintenance is Dulbecco's Modified Eagle's Medium (DMEM), but in a portion of published work another common medium, RPMI 1640, is substituted. RPMI 1640 lacks the high level of glucose present in DMEM (4.5 g/L) and it has been observed that the RAW 264.7 cells do not respond as well in this alternative environment. The purpose of this work compares RAW cells grown and maintained in DMEM and RPMI 1640 simultaneously. Both media are supplemented with 10% fetal bovine serum, sodium bicarbonate, and 100 IU Penicillin Streptomycin 100μg. DMEM is additionally made with 4.5 g/L glucose. Following a dose response exposure of RAW 264.7 to a range of 0 ng/mL to 100 ng/mL lipopolysaccharide from E. coli (serotype O55:B5) for 24 hours in a 5%CO2/ 37° C environment, nitrite produced by the cells is compared. Cells are kept in a continuous culture with passage every 3 days. Experimentation is carried out on a 24-well plate 500μl volume at 4x105 cells per well and incubated overnight. After exposure, Greiss reaction is utilized to measure nitric oxide response in collected supernatants. Initial results suggest differences in inflammatory responses, with DMEM appearing to elicit a more significant nitric oxide response. Once the difference between DMEM and RPMI 1640 has been established, a further goal of this work is to add glucose to RAW 264.7 that have undergone several passages in RPMI 1640 to determine if it is possible to raise their inflammatory response levels to what they would be in DMEM.
Faculty Mentor(s)
Dr. David Freier
Rights Statement
The right to download or print any portion of this material is granted by the copyright owner only for personal or educational use. The author/creator retains all proprietary rights, including copyright ownership. Any editing, other reproduction or other use of this material by any means requires the express written permission of the copyright owner. Except as provided above, or for any other use that is allowed by fair use (Title 17, §107 U.S.C.), you may not reproduce, republish, post, transmit or distribute any material from this web site in any physical or digital form without the permission of the copyright owner of the material.
Comparison of Two Standard Growth Media for the Inflammatory Response of Raw 264.7 Murine Macrophages
Turner Gymnasium
The RAW 264.7 murine macrophage is a significant cell line for the study of inflammation and one of the most extensively used cell lines in mammalian cell culture. The standard medium for its growth and maintenance is Dulbecco's Modified Eagle's Medium (DMEM), but in a portion of published work another common medium, RPMI 1640, is substituted. RPMI 1640 lacks the high level of glucose present in DMEM (4.5 g/L) and it has been observed that the RAW 264.7 cells do not respond as well in this alternative environment. The purpose of this work compares RAW cells grown and maintained in DMEM and RPMI 1640 simultaneously. Both media are supplemented with 10% fetal bovine serum, sodium bicarbonate, and 100 IU Penicillin Streptomycin 100μg. DMEM is additionally made with 4.5 g/L glucose. Following a dose response exposure of RAW 264.7 to a range of 0 ng/mL to 100 ng/mL lipopolysaccharide from E. coli (serotype O55:B5) for 24 hours in a 5%CO2/ 37° C environment, nitrite produced by the cells is compared. Cells are kept in a continuous culture with passage every 3 days. Experimentation is carried out on a 24-well plate 500μl volume at 4x105 cells per well and incubated overnight. After exposure, Greiss reaction is utilized to measure nitric oxide response in collected supernatants. Initial results suggest differences in inflammatory responses, with DMEM appearing to elicit a more significant nitric oxide response. Once the difference between DMEM and RPMI 1640 has been established, a further goal of this work is to add glucose to RAW 264.7 that have undergone several passages in RPMI 1640 to determine if it is possible to raise their inflammatory response levels to what they would be in DMEM.