Erica Castillo: Engineering the hiPSC-CM Microenvironment for cell-ECM Mechanobiology Studies
SEMINAR (Zoom)
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Speaker
Erica Castillo
9:00 am via zoom
Title: Engineering the hiPSC-CM Microenvironment for cell-ECM Mechanobiology Studies
Abstract
The biochemical and biophysical properties of the microenvironment are known to play a key role in cell structure and function. Specifically, the myocardium undergoes changes in extracellular matrix (ECM) proteins throughout cardiac development and disease. Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) hold great potential as a model to expand our knowledge of human heart muscle cells and their interactions with the surrounding microenvironment. Polyacrylamide (PA) hydrogels are a common mechanobiology substrate with a tunable physiological relevant stiffness range that also allows for functional quantitative measurements such as traction force microscopy (TFM). However, current protocols do not allow for the ECM protein type to be easily exchanged. Engineering microphysiological systems with tunable biochemical properties allow us to study the influence of ECM proteins on CM's ability to generate force against a load.
In this talk, I will discuss the development of our protocol which consists of a combination of lift-off protein patterning, a covalent protein-substrate linker, and copolymerization protein transfer. We show that we are able to tune the ECM protein type across a wide variety of proteins including Laminin, Fibronectin, Collagen IV, Collagen I, and Matrigel. And finally, I present TFM data comparing morphology and contraction force generation on Laminin versus Fibronectin, with Matrigel as a positive control. Lastly, I quantify the sarcomere organization, focal adhesions, and integrins among the different ECM proteins. Our developed protocol enables tuning microenvironment properties and quantifying cell-ECM interactions matched to cardiac development and disease states.
BIO
Dr. Erica Castillo is currently a Postdoctoral Scholar in the Mechanical Engineering Department with Prof. Beth Pruitt at UC Santa Barbara. Her current work focuses on developing microphysiological systems for heart muscle cell mechanobiology. Erica received her dual B.S. in Physics and Aerospace Engineering from the University of Texas at Arlington. In 2021, she received her Ph.D. in Mechanical Engineering from Stanford University with Prof. Beth Pruitt. Her graduate work developed a hydrogel-protein interface that allows heart muscle cells to interact with various ECM proteins. She also co-developed a wafer-scale protocol that increases the lithography throughput for hydrogel protein patterning. Erica’s work has been recognized with multiple awards and honors including an NSF Graduate Research Fellowship, a Ford Foundation Pre-Doctoral Fellowship, and a Stanford Mechanical Engineering Departmental Graduate Fellowship.
