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Isolation of Cardiomyocyte Nuclei from Post-mortem Tissue.

Bergmann, Olaf LU and Jovinge, Stefan LU (2012) In Journal of Visualized Experiments
Abstract
Identification of cardiomyocyte nuclei has been challenging in tissue sections as most strategies rely only on cytoplasmic marker proteins(1). Rare events in cardiac myocytes such as proliferation and apoptosis require an accurate identification of cardiac myocyte nuclei to analyze cellular renewal in homeostasis and in pathological conditions(2). Here, we provide a method to isolate cardiomyocyte nuclei from post mortem tissue by density sedimentation and immunolabeling with antibodies against pericentriolar material 1 (PCM-1) and subsequent flow cytometry sorting. This strategy allows a high throughput analysis and isolation with the advantage of working equally well on fresh tissue and frozen archival material. This makes it possible to... (More)
Identification of cardiomyocyte nuclei has been challenging in tissue sections as most strategies rely only on cytoplasmic marker proteins(1). Rare events in cardiac myocytes such as proliferation and apoptosis require an accurate identification of cardiac myocyte nuclei to analyze cellular renewal in homeostasis and in pathological conditions(2). Here, we provide a method to isolate cardiomyocyte nuclei from post mortem tissue by density sedimentation and immunolabeling with antibodies against pericentriolar material 1 (PCM-1) and subsequent flow cytometry sorting. This strategy allows a high throughput analysis and isolation with the advantage of working equally well on fresh tissue and frozen archival material. This makes it possible to study material already collected in biobanks. This technique is applicable and tested in a wide range of species and suitable for multiple downstream applications such as carbon-14 dating(3), cell-cycle analysis(4), visualization of thymidine analogues (e.g. BrdU and IdU)(4), transcriptome and epigenetic analysis. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Visualized Experiments
issue
65
publisher
JoVE
external identifiers
  • PMID:22805241
  • Scopus:84869819670
ISSN
1940-087X
DOI
10.3791/4205
language
English
LU publication?
yes
id
3b122999-72d7-4215-b3fb-25fcd1165530 (old id 2967025)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/22805241?dopt=Abstract
date added to LUP
2012-08-09 20:03:06
date last changed
2017-01-01 07:51:14
@article{3b122999-72d7-4215-b3fb-25fcd1165530,
  abstract     = {Identification of cardiomyocyte nuclei has been challenging in tissue sections as most strategies rely only on cytoplasmic marker proteins(1). Rare events in cardiac myocytes such as proliferation and apoptosis require an accurate identification of cardiac myocyte nuclei to analyze cellular renewal in homeostasis and in pathological conditions(2). Here, we provide a method to isolate cardiomyocyte nuclei from post mortem tissue by density sedimentation and immunolabeling with antibodies against pericentriolar material 1 (PCM-1) and subsequent flow cytometry sorting. This strategy allows a high throughput analysis and isolation with the advantage of working equally well on fresh tissue and frozen archival material. This makes it possible to study material already collected in biobanks. This technique is applicable and tested in a wide range of species and suitable for multiple downstream applications such as carbon-14 dating(3), cell-cycle analysis(4), visualization of thymidine analogues (e.g. BrdU and IdU)(4), transcriptome and epigenetic analysis.},
  author       = {Bergmann, Olaf and Jovinge, Stefan},
  issn         = {1940-087X},
  language     = {eng},
  number       = {65},
  publisher    = {JoVE},
  series       = {Journal of Visualized Experiments},
  title        = {Isolation of Cardiomyocyte Nuclei from Post-mortem Tissue.},
  url          = {http://dx.doi.org/10.3791/4205},
  year         = {2012},
}