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. 2019 Oct 10;9:14515. doi: 10.1038/s41598-019-51078-z

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Morphological evaluation of the in vitro interaction between telocytes (TCs) and satellite cells (SCs) from single living endomysial sheath-covered myofibers. (A,B) Phase-contrast microscopic images of endomysial sheath-covered myofibers from control and eccentric contraction (EC)-damaged muscles after 48 hours of culture on Matrigel-coated plates; sprouting cells appear more numerous around EC-injured myofibers. Note that some of these morphologically different cells display the typical features of TCs (i.e. moniliform/varicose telopodes; arrows and higher magnifications in the insets). (C,D) CD34 confocal immunofluorescence staining. Cells with a polygonal or spindle-shaped cell body abruptly giving rise to thin moniliform processes (arrows) and displaying a strong CD34 positivity, thus identifiable as TCs, are detected in the heterogeneous cell population derived from either control (C) or EC-injured (D) myofibers. These cells are distinguishable from typically fried egg-shaped CD34⁺ endothelial cells (D, inset). (E,F) WGA and Pax7 confocal double fluorescent staining. The presence of telopode-bearing TCs in control and EC-damaged myofiber-derived cells is confirmed by labeling with the membrane dye WGA. Pax7⁺ SCs, undetectable at this experimental time in the control myofiber-derived cell population (E), are instead detected in cell cultures from EC-damaged myofibers (F). Of note, SCs are often observed in cell pairs presumably resulting from mitotic division (F, inset) and in contact with TCs/telopodes (F, arrowhead and inset). Images are representative of at least 6 cell preparations from each of 3 control and 3 EC-injured muscle samples.