Human myoblasts from skeletal muscle biopsies: In vitro culture preparations for morphological and cytochemical analyses at light and electron microscopy

Manuela Malatesta, Marzia Giagnacovo, Rosanna Cardani, Giovanni Meola, Carlo Pellicciari

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

We describe protocols for the isolation of satellite cells from human muscle biopsies, for the in vitro culture of proliferating and differentiating myoblasts, and for the preparation of cell samples suitable for morphological and cytochemical analyses at light and electron microscopy. The procedures described are especially appropriate for processing small muscle biopsies, and allow obtaining myoblast/myotube monolayers on glass coverslips, thus preserving good cell morphology and immunoreactivity for protein markers of myoblast proliferation, differentiation, and senescence. These cell preparations are suitable for cytochemical, immunocytochemical, and FISH procedures at light microscopy, and can be observed not only in brightfield, phase contrast, and differential interference contrast but also in fluorescence (which can hardly be used for cells grown on conventional plastic surfaces, which generally exhibit intense auto fluorescence). In their ultrastructural cytochemical application, the protocols are intended for post-embedding techniques, by which ultrathin sections from a single sample may be used for detecting a wide variety of molecular markers.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages67-79
Number of pages13
Volume976
ISBN (Print)9781627033169
DOIs
Publication statusPublished - 2013

Publication series

NameMethods in Molecular Biology
Volume976
ISSN (Print)10643745

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Keywords

  • Cytochemistry
  • In vitro differentiation
  • In vitro proliferation
  • In vitro senescence
  • Light microscopy
  • Morphology
  • Myoblasts
  • Transmission electron microscopy

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Medicine(all)

Cite this

Malatesta, M., Giagnacovo, M., Cardani, R., Meola, G., & Pellicciari, C. (2013). Human myoblasts from skeletal muscle biopsies: In vitro culture preparations for morphological and cytochemical analyses at light and electron microscopy. In Methods in Molecular Biology (Vol. 976, pp. 67-79). (Methods in Molecular Biology; Vol. 976). Humana Press Inc.. https://doi.org/10.1007/978-1-62703-317-6_6