Establishment of Primary Neuronal Cultures for the Investigation of Neuronal Survival In Vitro
Location
CSU Ballroom
Start Date
21-4-2014 10:00 AM
End Date
21-4-2014 11:30 AM
Student's Major
Biological Sciences
Student's College
Science, Engineering and Technology
Mentor's Name
Rachel Bergstrom
Mentor's Department
Biological Sciences
Mentor's College
Science, Engineering and Technology
Description
Neurons are cells of the brain that pose a challenge to study in vitro as they are one of the few cell types that are post-mitotic and cannot be maintained for a tissue culture as a cell line. Therefore, they must be taken from live organisms and grown in a culture. This study used mouse pups at embryonic day fifteen. A standard procedure for setting up neuron cultures for testing was developed and used for future experiments. Extreme care was taken to prevent cultured neurons from coming in contact with contaminating substances, such as bacteria, by handling neurons in a sterile tissue culture hood. In vitro, neurons require a growth media with all nutrients required for growth and survival. One specific nutrient mixture is B27, which is a standard nutrient mixture of 27 components for neuronal growth in vitro. To study pro-survival protein signaling events within the neurons, serum starve time courses were completed, depriving the neurons from B27 for different lengths of time, including a control group which was not deprived of B27. Proteins were then extracted through cell lysis and analyzed against the control with SDS gel electrophoresis. The comparison of protein expression between serum starved and not starved neurons will allow us to begin to understand what is involved in the pro-survival signaling pathways of neurons. Culturing neurons in vitro is vital to understanding the cellular and molecular components of neuron function.
Establishment of Primary Neuronal Cultures for the Investigation of Neuronal Survival In Vitro
CSU Ballroom
Neurons are cells of the brain that pose a challenge to study in vitro as they are one of the few cell types that are post-mitotic and cannot be maintained for a tissue culture as a cell line. Therefore, they must be taken from live organisms and grown in a culture. This study used mouse pups at embryonic day fifteen. A standard procedure for setting up neuron cultures for testing was developed and used for future experiments. Extreme care was taken to prevent cultured neurons from coming in contact with contaminating substances, such as bacteria, by handling neurons in a sterile tissue culture hood. In vitro, neurons require a growth media with all nutrients required for growth and survival. One specific nutrient mixture is B27, which is a standard nutrient mixture of 27 components for neuronal growth in vitro. To study pro-survival protein signaling events within the neurons, serum starve time courses were completed, depriving the neurons from B27 for different lengths of time, including a control group which was not deprived of B27. Proteins were then extracted through cell lysis and analyzed against the control with SDS gel electrophoresis. The comparison of protein expression between serum starved and not starved neurons will allow us to begin to understand what is involved in the pro-survival signaling pathways of neurons. Culturing neurons in vitro is vital to understanding the cellular and molecular components of neuron function.
Recommended Citation
Hanson, Taylour and Paul Creger. "Establishment of Primary Neuronal Cultures for the Investigation of Neuronal Survival In Vitro." Undergraduate Research Symposium, Mankato, MN, April 21, 2014.
https://cornerstone.lib.mnsu.edu/urs/2014/poster_session_A/13
