Mentors
Project Description
Cardiomyocyte (CM) culture is a useful platform to model cardiac biology and disease. However, iPSC-derived CMs in 2D often have heterogeneous characteristics both within and between differentiations. This is, in part, due to the relatively immature state of these cells. Most notably, these cells lack uniform and aligned sarcomere structures. Using microfabrication techniques, this project aims to optimize a 2D CM culture device that promotes sarcomere alignment and uniformity.
This will enable the functional characterization of CMs derived from isogenic iPSC cell lines harboring various sarcomere mutations in order to better understand how these mutations cause disease. These devices could also serve as a platform to test therapies for said mutations. Specifically, this project will develop a protein-micropattern fabrication protocol for use in CM culture.
Additionally, the substrate stiffness of this culture platform can be experimentally modified to optimize substrate elasticity for sarcomere development. Sarcomere assessment will then be employed in various culture conditions via microscopy and computational analysis.
Research Participant
Program: CELL-MET REU
Sharvari Apte, Research Experiences for Undergraduates.
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Research Goals
- Fabricate culture substrates with a variety of protein micropatterns via microcontact printing.
- Experimentally control PDMS culture substrate stiffness, quantify substrate stiffness.
- Study the impact of micropattern geometry/substrate stiffness on sarcomere development – optimize conditions for sarcomere development.
- Characterize morphological and functional differences between wildtype and mutant CMs.
Learning Goals
- Through this project, the student will develop technical skills related to cardiac tissue engineering as well as more general scientific skills such as iterative design and hypothesis development.
- Specifically, this student will learn stem cell maintenance and differentiation, microfabrication techniques, PDMS device fabrication, microscopy, immunofluorescence, and functional cardiomyocyte assessment via computational analysis techniques.
Timeline
Weeks 1-2: Technical skills building/onboarding.
Weeks 3-5: Building familiarity with the skills learned through pilot experimentation.
Weeks 6-9: Iterative optimization of device and finish experimentation.
Week 10: Finish up data analysis and synthesize final presentation.
