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Embryonic stem cells in predictive cardiotoxicity: laser capture microscopy enables assay development.
Khuram W Chaudhary1, Nestor X Barrezueta, Mary B Bauchmann
1Pfizer Global Research and Development, Chesterfield, Missouri 63017, USA.
Summary
Laser microdissection and pressure catapulting (LMPC) isolates pure embryonic stem (ES) cell-derived cardiomyocytes. This method enables reliable in vitro drug discovery and cardiotoxicity testing for safer novel drugs.
Area of Science:
- Stem cell biology
- Cardiovascular research
- Drug discovery
Background:
- Embryonic stem (ES) cells are valuable for in vitro drug discovery and safety testing.
- ES cell-derived cardiomyocytes can predict drug cardiotoxicity.
- Cellular heterogeneity in differentiated ES cells hinders reliable in vitro model development.
Purpose of the Study:
- To develop a robust method for isolating homogeneous ES cell-derived cardiomyocytes.
- To enable reliable in vitro pharmacology assays for drug screening and cardiotoxicity assessment.
Main Methods:
- Utilized laser microdissection and pressure catapulting (LMPC) to isolate ES-derived cardiomyocytes.
- Applied isolated cells to 96-well format in vitro pharmacology assays.
- Confirmed expression of key cardiac developmental and functional markers (Nkx 2.5, MLC2V, GATA-4, Connexin 43, Connexin 45, Serca-2a, cardiac alpha actin, phospholamban).
Main Results:
- LMPC successfully isolated homogeneous ES-derived cardiomyocytes.
- Isolated cells expressed essential cardiac markers.
- Functional assays demonstrated expected cardiac responses to calcium and nifedipine.
Conclusions:
- Laser microdissection and pressure catapulting (LMPC) is a robust technology for isolating homogeneous ES-derived cell types.
- This method is applicable to developing reliable in vitro assays for drug discovery and cardiotoxicity testing.