Location

Schewel Hall Room 222

Access Type

Campus Access Only

Presentation Type

Oral Presentation

Event Website

http://www.lynchburg.edu/academics/red-letter-day/student-scholar-showcase/

Start Date

6-4-2016 9:30 AM

End Date

6-4-2016 9:45 AM

Abstract

Culture wells exist in various sizes determined by the number of wells on a plate and the shape of the well bottoms. The possible effect of culture well dimensions on the cells within is often ignored. The goal of this experiment was to examine the physical dynamics of the culture environment as a 3-dimensional system using round and flat-bottomed 96-well culture plates. Nitric oxide (NO) production from RAW 264.7 murine macrophages stimulated by lipopolysaccharide (LPS) from E. coli (O55:B5) was used as a model system to observe the effect of culture well shape on a well-characterized biological phenomenon. Replicable conditions for macrophage NO response were established for the 96-well culture plates, all experiments being done in 200 μL well volumes. It was hypothesized that round-bottomed wells offered greater surface area for cell adherence, leading to a higher NO response. LPS treatments used to create the dose-response curve of 0, 0.1, 1, 10, and 100 ng/mL respectively were tested based upon previous work. A range of cell densities from 2x104 to 4x105 cells/well was chosen for testing based upon values commonly seen in literature. No difference in NO levels following LPS stimulation based upon cell density and well shape were observed.

Faculty Mentor

Dr. David O. Freier

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Apr 6th, 9:30 AM Apr 6th, 9:45 AM

The effect of culture well surface topography on nitric oxide production of E. coli LPS-stimulated RAW 264.7 murine macrophages

Schewel Hall Room 222

Culture wells exist in various sizes determined by the number of wells on a plate and the shape of the well bottoms. The possible effect of culture well dimensions on the cells within is often ignored. The goal of this experiment was to examine the physical dynamics of the culture environment as a 3-dimensional system using round and flat-bottomed 96-well culture plates. Nitric oxide (NO) production from RAW 264.7 murine macrophages stimulated by lipopolysaccharide (LPS) from E. coli (O55:B5) was used as a model system to observe the effect of culture well shape on a well-characterized biological phenomenon. Replicable conditions for macrophage NO response were established for the 96-well culture plates, all experiments being done in 200 μL well volumes. It was hypothesized that round-bottomed wells offered greater surface area for cell adherence, leading to a higher NO response. LPS treatments used to create the dose-response curve of 0, 0.1, 1, 10, and 100 ng/mL respectively were tested based upon previous work. A range of cell densities from 2x104 to 4x105 cells/well was chosen for testing based upon values commonly seen in literature. No difference in NO levels following LPS stimulation based upon cell density and well shape were observed.

https://digitalshowcase.lynchburg.edu/studentshowcase/2016/Presentations/12