Micropattern width dependent sarcomere development in human ESC-derived cardiomyocytes

Max R. Salick, Brett N. Napiwocki, Jin Sha, Gavin T. Knight, Shahzad A. Chindhy, Timothy J. Kamp, Randolph S. Ashton, Wendy C. Crone

Research output: Contribution to journalArticlepeer-review

125 Scopus citations


In this study, human embryonic stem cell-derived cardiomyocytes were seeded onto controlled two-dimensional micropatterned features, and an improvement in sarcomere formation and cell alignment was observed in specific feature geometries. High-resolution photolithography techniques and microcontact printing were utilized to produce features of various rectangular geometries, with areas ranging from 2500μm2 to 160,000μm2. The microcontact printing method was used to pattern non-adherent poly(ethylene glycol) regions on gold coated glass slides. Matrigel and fibronectin extracellular matrix (ECM) proteins were layered onto the gold-coated glass slides, providing a controlled geometry for cell adhesion. We used small molecule-based differentiation and an antibiotic purification step to produce a pure population of immature cardiomyocytes from H9 human embryonic stem cells (hESCs). We then seeded this pure population of human cardiomyocytes onto the micropatterned features of various sizes and observed how the cardiomyocytes remodeled their myofilament structure in response to the feature geometries. Immunofluorescence was used to measure α-actinin expression, and phalloidin stains were used to detect actin presence in the patterned cells. Analysis of nuclear alignment was also used to determine how cell direction was influenced by the features. The seeded cells showed clear alignment with the features, dependent on the width rather than the overall aspect ratio of the features. It was determined that features with widths between 30μm and 80μm promoted highly aligned cardiomyocytes with a dramatic increase in sarcomere alignment relative to the long axis of the pattern. This creation of highly-aligned cell aggregates with robust sarcomere structures holds great potential in advancing cell-based pharmacological studies, and will help researchers to understand the means by which ECM geometries can affect myofilament structure and maturation in hESC-derived cardiomyocytes.

Original languageEnglish (US)
Pages (from-to)4454-4464
Number of pages11
Issue number15
StatePublished - May 2014
Externally publishedYes

Bibliographical note

Funding Information:
This research was supported with funds from the National Institutes of Health Grant K18 HL105504 from the Heart, Blood and Lung Institute and the Graduate School of the University of Wisconsin-Madison .

Copyright 2014 Elsevier B.V., All rights reserved.


  • Cardiac tissue engineering
  • Cardiomyocyte
  • Cell morphology
  • Micropatterning
  • Stem cell
  • Surface modification


Dive into the research topics of 'Micropattern width dependent sarcomere development in human ESC-derived cardiomyocytes'. Together they form a unique fingerprint.

Cite this