Assessment of Cell Viability in Electrically Excitable Muscle Cells Through Intact Twitch Stimulation

Bauer J, Head SI, Friedrich O (2023)

Publication Type: Book chapter / Article in edited volumes

Publication year: 2023

Journal

Methods in Molecular Biology Humana Press

Publisher: Humana Press Inc.

Edited Volumes: Cell Viability Assays (2nd Edt.)

Series: Methods in Molecular Biology

Book Volume: 2644

Pages Range: 177-192

DOI: 10.1007/978-1-0716-3052-5_11

Abstract

Muscle cells (i.e. skeletal muscle fibers) are fully viable and functional when their excitation-contraction (EC) coupling machinery is intact. This involves intact membrane integrity with polarized membrane, functional ion channels for action potential generation and conduction, an intact electro-chemical interface at the level of the fiber’s triad, followed by sarcoplasmic reticulum Ca2+ release, and subsequent activation of the chemico-mechanical interface at the level of the contractile apparatus. The ultimate end result is then a visible twitch contraction upon a brief electrical pulse stimulation. For many biomedical studies involving single muscle cells, intact and viable myofibers are of utmost importance. Thus, a simple global screening method that involves a brief electrical stimulus applied to single muscle fibers and assessment of visible contraction would be of high value. In this chapter, we describe step-by-step protocols to (i) obtain intact single muscle fibers from freshly dissected muscle tissue using an enzymatic digestion procedure and (ii) provide a workflow for the assessment of twitch response of single fibers that can be ultimately classified as viable. For this, we have prepared a unique stimulation pen for which we provide the fabrication guide for do-it-yourself rapid prototyping to eliminate the need for expensive specialized commercial equipment.

Authors with CRIS profile

Julian Bauer Lehrstuhl für Medizinische Biotechnologie (MBT) Oliver Friedrich Lehrstuhl für Medizinische Biotechnologie (MBT)

Involved external institutions

University of Western Sydney AU Australia (AU)

How to cite

APA:

Bauer, J., Head, S.I., & Friedrich, O. (2023). Assessment of Cell Viability in Electrically Excitable Muscle Cells Through Intact Twitch Stimulation. In Friedrich O, Gilbert DF (Eds.), Cell Viability Assays (2nd Edt.). (pp. 177-192). Humana Press Inc..

MLA:

Bauer, Julian, Stewart I. Head, and Oliver Friedrich. "Assessment of Cell Viability in Electrically Excitable Muscle Cells Through Intact Twitch Stimulation." Cell Viability Assays (2nd Edt.). Ed. Friedrich O, Gilbert DF, Humana Press Inc., 2023. 177-192.

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