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
Marilyn Hart
Mentor's Email Address
marilyn.hart@mnsu.edu
Mentor's Department
Biological Sciences
Mentor's College
Science, Engineering and Technology
Second Mentor's Name
Geoffrey Goellner
Second Mentor's Email Address
geoffrey.goellner@mnsu.edu
Second Mentor's Department
Biological Sciences
Second Mentor's College
Science, Engineering and Technology
Third Mentor's Name
Michael Bentley
Third Mentor's Email Address
michael.bentley@mnsu.edu
Third Mentor's Deparment
Biological Sciences
Third Mentor's College
Science, Engineering and Technology
Description
Alterations of sarcomeric proteins lead to disruption of myofilaments and are associated with hypertrophic cardiomyopathy. We have identified a genetically altered mouse strain with an elevated level of actin associated protein and are characterizing the nature of the hypertrophy by examining the cultured cells on glass microcarrier beads using Scanning Electron Microscopy (SEM). Beads provide a surface for cell growth and division and subsequent analysis of myocyte morphology. This study requires the establishment of primary embryonic cardiomyocyte culture which is difficult to establish. Therefore in initial studies to acquire the necessary tissue culture skill, we cultured Human Embryonic Kidney (HEK) cells. Confluent HEK cell cultures were established and the cells used to plate collagen coated dextran microcarrier beads (60-87µm) using varying bead concentrations. The cells were plated at low density, incubated at 37°C for four days in the presence of 5% CO2. The cells, attached to the microcarrier beads, were preserved by fixation in 2.5% glutaraldehyde and visualized using SEM. The shape, size, and filopodia of the HEK cells were characterized, demonstrating the feasibility of this technique. We are currently establishing primary cell cultures of mouse embryonic cardiomyocytes, from both wild type and genetically altered mice with known sarcomeric disarray. The individual myocytes will be analyzed for alterations at the cellular level.
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Included in
Examination of Human Embryonic Kidney cells and Cardiomyocytes using Glass Microcarrier Beads and Scanning Electron Microscopy
CSU Ballroom
Alterations of sarcomeric proteins lead to disruption of myofilaments and are associated with hypertrophic cardiomyopathy. We have identified a genetically altered mouse strain with an elevated level of actin associated protein and are characterizing the nature of the hypertrophy by examining the cultured cells on glass microcarrier beads using Scanning Electron Microscopy (SEM). Beads provide a surface for cell growth and division and subsequent analysis of myocyte morphology. This study requires the establishment of primary embryonic cardiomyocyte culture which is difficult to establish. Therefore in initial studies to acquire the necessary tissue culture skill, we cultured Human Embryonic Kidney (HEK) cells. Confluent HEK cell cultures were established and the cells used to plate collagen coated dextran microcarrier beads (60-87µm) using varying bead concentrations. The cells were plated at low density, incubated at 37°C for four days in the presence of 5% CO2. The cells, attached to the microcarrier beads, were preserved by fixation in 2.5% glutaraldehyde and visualized using SEM. The shape, size, and filopodia of the HEK cells were characterized, demonstrating the feasibility of this technique. We are currently establishing primary cell cultures of mouse embryonic cardiomyocytes, from both wild type and genetically altered mice with known sarcomeric disarray. The individual myocytes will be analyzed for alterations at the cellular level.
Recommended Citation
Sim, Jaekook. "Examination of Human Embryonic Kidney cells and Cardiomyocytes using Glass Microcarrier Beads and Scanning Electron Microscopy." Undergraduate Research Symposium, Mankato, MN, April 21, 2014.
https://cornerstone.lib.mnsu.edu/urs/2014/poster_session_A/22
