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 language | English |
|---|---|
| Title of host publication | Methods in Molecular Biology |
| Publisher | Humana Press Inc. |
| Pages | 67-79 |
| Number of pages | 13 |
| Volume | 976 |
| ISBN (Print) | 9781627033169 |
| DOIs | |
| Publication status | Published - 2013 |
Publication series
| Name | Methods in Molecular Biology |
|---|---|
| Volume | 976 |
| 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
Human myoblasts from skeletal muscle biopsies : In vitro culture preparations for morphological and cytochemical analyses at light and electron microscopy. / Malatesta, Manuela; Giagnacovo, Marzia; Cardani, Rosanna; Meola, Giovanni; Pellicciari, Carlo.
Methods in Molecular Biology. Vol. 976 Humana Press Inc., 2013. p. 67-79 (Methods in Molecular Biology; Vol. 976).Research output: Chapter in Book/Report/Conference proceeding › Chapter
}
TY - CHAP
T1 - Human myoblasts from skeletal muscle biopsies
T2 - In vitro culture preparations for morphological and cytochemical analyses at light and electron microscopy
AU - Malatesta, Manuela
AU - Giagnacovo, Marzia
AU - Cardani, Rosanna
AU - Meola, Giovanni
AU - Pellicciari, Carlo
PY - 2013
Y1 - 2013
N2 - 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.
AB - 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.
KW - Cytochemistry
KW - In vitro differentiation
KW - In vitro proliferation
KW - In vitro senescence
KW - Light microscopy
KW - Morphology
KW - Myoblasts
KW - Transmission electron microscopy
UR - http://www.scopus.com/inward/record.url?scp=84880620420&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84880620420&partnerID=8YFLogxK
U2 - 10.1007/978-1-62703-317-6_6
DO - 10.1007/978-1-62703-317-6_6
M3 - Chapter
SN - 9781627033169
VL - 976
T3 - Methods in Molecular Biology
SP - 67
EP - 79
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -