InsideAnswers: Techniques and Best Practices for Cardiomyocyte Isolation
In this Q&A video series sponsored by IonOptix, experts address your best questions and greatest challenges with regards to cardiomyocyte isolation, purification, culture and analysis. These questions have been adapted from registrants’ input during our November 2018 webinar, Techniques and Best Practices for Cardiomyocyte Isolation.
Presenters
Professor of Medicine
Cardiac Muscle Research Laboratory
Stanford University School of Medicine
rliao7@stanford.edu
Lecturer of Cardiovascular Sciences
College of Medical and Dental Sciences
University of Birmingham
D.Pavlovic@bham.ac.uk
Post Doctoral Fellow
Yong Loo Lin School of Medicine
National University of Singapore
mdcmaa@nus.edu.sg
Discussion Topics:
- Strategies to consistently improve your isolation success
- Tips for getting started with the Langendorff-free method of cardiomyocyte isolation
- Ideal short- and long-term culture conditions
- Which enzymes to use and how to ensure the heart is optimally digested
Associated Resources:
- Ackers-Johnson, M. et al (2016). A Simplified, Langendorff-Free Method for Concomitant Isolation of Viable Cardiac Myocytes and Nonmyocytes From the Adult Mouse Heart
- Liao Lab Protocol: Rat Cardiomyocyte Isolation Using non-Langendorff Mode
- Liao Lab Protocol: Rat Cardiomyocyte Isolation Using Langendorff Mode
- Liao Lab Protocol: Adult Mouse Cardiomyocyte Culture Procedure
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How long should it take to excise the heart and hang it on the Langendorff system?
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What are some strategies to help improve cardiomyocyte isolation skill?
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What portion of isolated cardiomoyocytes respond to pacing?
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When isolating cardiomyocytes, how should perfusion speed be adjusted based on mouse or rat weight?
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What are some considerations when using cardiomyocytes 2–3 days after isolation?
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How important is temperature for successful cardiomyocyte isolation and analysis?
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General tips and user feedback using the Langendorff-free method for cardiomyocyte isolation
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BDM and cardiomyocyte isolation: when should you use it and when is it best avoided?
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How to purify cardiomyocytes
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Tips for short- and long-term culturing of adult mouse cardiomyocytes
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If you do not have much experience with cannulation, should you use the injection method?
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How to transfer cardiomyocytes from Ca2+-free to Ca2+ buffer
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When using the Injection method of cardiomyocyte isolation, is rupturing the heart a common problem?
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Which enzyme (or mixture of enzymes) is best for digestion of the heart for cardiomyocyte isolation?
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When digesting the heart for cardiomyocyte isolation, how long does it usually take?
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