Analysis and quantification of in vitro myoblast fusion using the LADD Multiple Stain

作者：McColl Rhys; Nkosi Mthokozisi; Snyman Celia; Niesler Carola^*

来源：Biotechniques, 2016, 61(6): 323-326.

DOI：10.2144/000114485

• fusion
• myoblast
• microscopy
• LADD
• skeletal muscle
• differentiation

Myoblast fusion, which is essential for muscle development, regeneration, and repair, can be assessed in vitro via the calculation of a fusion index. Traditionally, this requires use of either immunocytochemistry or fluorescently-labeled cytoskeletal staining, followed by microscopy and laborious analysis. The expense and time-consuming nature of the optimization and application of antibody-based techniques such as immunocytochemistry, as well as the need for specialized analytical equipment such as fluorescence microscopes, presents a barrier to the routine analysis of this crucial step during terminal differentiation. Here, we describe (i) a novel use of the commonly available LADD Multiple Stain for visualization of myoblast fusion in vitro; (ii) the optimization of a simple image analysis method to generate quick, quantifiable data representative of a fusion index; and (M) the use of a protocol combining these two procedures to investigate in vitro myoblast fusion in a simple and efficient manner as proof-of-concept.

• 出版日期2016-12