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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Creative Commons Attribution 4.0 License
This work is licensed under a Creative Commons Attribution 4.0 License.

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Apr 21st, 10:00 AM Apr 21st, 11:30 AM

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.
http://cornerstone.lib.mnsu.edu/urs/2014/poster_session_A/22

 

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