Non Animal Testing Database

Software tool for automated assessment of sarcomeres in muscle cell cultures

Leiden University Medical Center, Leiden, Netherlands
Sarcomeres are the structural units of the contractile apparatus in cardiac and skeletal muscle cells. Assessment of sarcomere length, alignment, and organization provides insight into disease and drug responses in striated muscle cells and models, ranging from cardiomyocytes and skeletal muscle cells derived from human pluripotent stem cells to adult muscle cells isolated from humans. However, quantification of sarcomere length is typically time-consuming and prone to user-specific selection bias. Automated analysis pipelines exist but these often require either specialized software or programming experience. In addition, these pipelines are often designed for only one type of cell model in vitro. Here, an easy-to-implement protocol and software tool for automated sarcomere length and organization quantification in a variety of striated muscle in vitro models was presented: Two-dimensional (2D) cardiomyocytes, three-dimensional (3D) cardiac microtissues, isolated adult cardiomyocytes, and 3D tissue-engineered skeletal muscles. Based on an existing mathematical algorithm, this image analysis software (SotaTool) automatically detects the direction in which the sarcomere organization is highest over the entire image and outputs the length and organization of sarcomeres. Videos of live cells during contraction were also analysed, thereby allowing the measurement of contraction parameters like fractional shortening, contraction time, relaxation time, and beating frequency. This protocol gives a step-by-step guide on how to prepare, image, and automatically quantify sarcomere and contraction characteristics in different types of in vitro models and provides basic validation and discussion of the limitations of the software tool.
Software tool for automatic quantification of sarcomere length and organization in fixed and live 2D and 3D muscle cell cultures In vitro
Berend J. Van Meer
Added on: 12-14-2022
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