Mary Gates Endowment For Students

Alan Levinson

Mary Gates Research Scholar, Winter 2021 & Winter 2022

 Research Project: Engineered Heart Tissue Image Processing Suite

Project Description: Induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) that have been engineered into three-dimensional heart tissues (EHTs) are valuable research tools for investigating debilitating genetic diseases that afflict the heart, such as Duchenne muscular dystrophy (DMD). Ensuring iPSC-CMs can be sufficiently matured to model such diseases remains a hurdle in current research, and maturational analysis techniques for iPSC-CMs are either qualitative, manual, or primarily based in two dimensions, leaving much to be desired. This poster details the creation of a suite of MATLAB image-processing scripts that can quantify the effect of three-dimensional culture and disease-causing DMD mutations on cardiomyocyte structure and maturation state. The iPSC-CMs were differentiated from stem cells, cast into EHTs, stained using immunofluorescence, and imaged using confocal microscopy. Using the scripts to analyze these 3D images of iPSC-CM stains, key maturational features of the cells can be quantified such as nuclei count; cardiomyocyte area; and sarcomere length, orientation, and z-disk width. Analyzing cardiomyocyte area can give key information on cardiomyocyte hypertrophy while examining sarcomere length, orientation, and Z-disk width can provide information on myofibril structure and organization. The suite allows analysis of these maturational features in both 2D and 3D cultures and offers a method for quantitatively assessing maturation in an automated manner. Measuring iPSC-CM maturation will also allow better comparison of existing maturational methods, such as mechanical loading, electrical stimulation, and small molecule treatment. The suite can also create graphical outputs to elegantly display data. Recent progress also includes a script that can count cell nuclei and quantify cell area. Overall, the suite will help improve maturational analysis of EHTs, and hopefully contribute to the discovery of new treatments for diseases that affect the heart. 

What have you learned throughout your research project?

I have learned the value of presenting and getting feedback on research in order to make the stories that research tells more coherent and impactful. I also learned many wet lab research skills as well as coding skills that I hope to use extensively later in life.

What piece of advice do you have for future applicants?

Don’t be afraid to reach out to mentors for suggestions and advice to help guide your research. Also, take advantage of opportunities to share your research with friends and other researchers.