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Fibroblast Derived Human Engineered Connective Tissue for Screening Applications
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1Institute of Pharmacology and Toxicology,University Medical Center Goettingen, 2DZHK (German Center for Cardiovascular Research) partner site, Goettingen, 3Cluster of Excellence “Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells” (MBExC),University of Goettingen, 4Center for Neurodegenerative Diseases (DZNE), 5Fraunhofer Institute for Translational Medicine and Pharmacology (ITMP)
Chapters
- 00:04Introduction
- 00:52Primary Cardiac Fibroblast Preparation
- 01:39Enzymatic Dispersion of Human Cardiac Fibroblasts
- 02:52Engineered Connective Tissue Preparation
- 05:28Cross-Sectional Area Measurement to Assess ECT Compaction
- 05:55Pole Deflection Analysis to Monitor ECT Contraction
- 06:32Destructive Tensile Measurement and Stress-Strain Analysis to Assess ECT
- 08:20Results: Effect of Actin Polymerization on ECT Compaction, Contraction, and Stiffness
- 09:22Conclusion
Presented here is a protocol to generate engineered connective tissues for a parallel culture of 48 tissues in a multi-well plate with double poles, suitable for mechanistic studies, disease modeling, and screening applications. The protocol is compatible with fibroblasts from different organs and species and is exemplified here with human primary cardiac fibroblasts.
