ArticleCell biochemistry and biophysics2011
An improved isolation procedure for adult mouse cardiomyocytes.
Article in Cell biochemistry and biophysics, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 26 citations in OpenAlex.
- Mitochondrial connexin43 and mitochondrial KPflugers Archiv : European journal of physiology · 2023Article
- Isolation, Transfection, and Long-Term Culture of Adult Mouse and Rat Cardiomyocytes.Journal of visualized experiments : JoVE · 2020Article
- Simultaneous Isolation and Culture of Atrial Myocytes, Ventricular Myocytes, and Non-Myocytes from an Adult Mouse Heart.Journal of visualized experiments : JoVE · 2020Article
- Proteomic analysis of the cardiac myocyte secretome reveals extracellular protective functions for the ER stress response.Journal of molecular and cellular cardiology · 2020Article
- Many Cells Make Life Work-Multicellularity in Stem Cell-Based Cardiac Disease Modelling.International journal of molecular sciences · 2018Review
- Fibroblast growth factor-21 prevents diabetic cardiomyopathy via AMPK-mediated antioxidation and lipid-lowering effects in the heart.Cell death & disease · 2018Article
- A Simplified, Langendorff-Free Method for Concomitant Isolation of Viable Cardiac Myocytes and Nonmyocytes From the Adult Mouse Heart.Circulation research · 2016Article
- Isolation, Culture and Transduction of Adult Mouse Cardiomyocytes.Journal of visualized experiments : JoVE · 2016Article
- Heterozygous caveolin-3 mice show increased susceptibility to palmitate-induced insulin resistance.Physiological reports · 2016Article
- In Vivo and In Silico Investigation Into Mechanisms of Frequency Dependence of Repolarization Alternans in Human Ventricular Cardiomyocytes.Circulation research · 2016Article
- Article
- Live cell imaging of primary rat neonatal cardiomyocytes following adenoviral and lentiviral transduction using confocal spinning disk microscopy.Journal of visualized experiments : JoVE · 2014Article
- Palmitate diet-induced loss of cardiac caveolin-3: a novel mechanism for lipid-induced contractile dysfunction.PloS one · 2013Article
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Authors and funding
4 authors at 3 institutions in 1 country.
Funding
Abstract
Isolated adult mouse cardiomyocytes are an important tool in cardiovascular research, but are challenging to prepare. Because the energy supply determines cell function and viability, we compared total creatine ([Cr]) and [ATP] in isolated cardiomyocytes with the intact mouse heart. Isolated myocytes suffered severe losses of Cr (-70%) and ATP (-53%). Myocytes were not able to replete [Cr] during a 5 h incubation period in medium supplemented with 1 mM Cr. In contrast, adding 20 mM Cr to the digestion buffers was sufficient to maintain normal [Cr]. Supplementing buffers with 5 mM of inosine (Ino) and adenosine (Ado) to prevent loss of cellular nucleosides partially protected against loss of ATP. To test whether maintaining [ATP] and [Cr] improves contractile function, myocytes were challenged by varying pacing rate from 0.5 to 10 Hz and by adding isoproterenol (Iso) at 5 and 10 Hz. All groups performed well up to 5 Hz, showing a positive cell shortening-frequency relationship; however, only 16% of myocytes isolated under standard conditions were able to sustain pacing with Iso challenge at 10 Hz. In contrast, 30-50% of the myocytes with normal Cr levels were able to contract and maintain low diastolic [Ca(2+)]. Cell yield also improved in Cr and the Cr/Ino/Ado-treated groups (85-90% vs. 70-75% rod shaped in untreated myocytes). These data suggest that viability and performance of isolated myocytes are improved when they are protected from the severe loss of Cr and ATP during the isolation, making them an even better research tool.
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