By Milica Radisic, Lauren D. Black III
Cardiac Tissue Engineering: tools and Protocols presents a set of protocols on cardiac tissue engineering from pioneering and major researchers world wide. those comprise equipment and protocols for mobilephone practise, biomaterial coaching, cellphone seeding, and cultivation in a variety of structures. Written within the hugely profitable Methods in Molecular Biology sequence structure, chapters comprise introductions to their respective subject matters, lists of the required fabrics and reagents, step by step, quite simply reproducible laboratory protocols, and key pointers on troubleshooting and heading off recognized pitfalls.
Authoritative and sensible, Cardiac Tissue Engineering: tools and Protocols highlights the main innovations, either experimental and computational, for the examine of cardiovascular tissue engineering.
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Additional resources for Cardiac Tissue Engineering: Methods and Protocols
Culture until the clonal candidates reach 80 % confluency in the T75 culture flasks, 1–2 weeks. Exchange media every 7 days. 27. The cells are now ready for characterization and further expansion of c-kit-positive clones. 28. Trypsinize and pellet the cells. 29. Resuspend the cells in culture media and count. 30. Set aside 1 × 105 cells per desired stain (at least 5 × 105 cells) for characterization. 31. Seed the remaining cells at 5 × 105 to 1 × 106 cells per T75 culture flask (see Note 13). 32.
Khan SS, Solomon MA, McCoy JP Jr (2005) Detection of circulating endothelial cells and endothelial progenitor cells by flow cytometry. Cytometry B Clin Cytom 64:1–8 28. Shepherd RM, Capoccia BJ, Devine SM et al (2006) Angiogenic cells can be rapidly mobilized and efficiently harvested from the blood following treatment with AMD3100. Blood 108:3662–3667 29. Steinmetz M, Nickenig G, Werner N (2010) Endothelial-regenerating cells: an expanding universe. Hypertension 55:593–599 30. Yip HK, Chang LT, Chang WN et al (2008) Level and value of circulating endothelial progenitor cells in patients after acute ischemic stroke.
Bauwens and Mark D. Ungrin 2. Use a 5 mL serological pipette to transfer the aggregates to 15 mL conical tubes, pooling up to 10 mL of aggregates per tube. Allow 10 min for the aggregates to settle. Aspirate the supernatant and resuspend the aggregates in Stage 3 Induction Medium. Using a 5 mL serological pipette, redistribute the aggregates into a 24 well ULA plate at 1 mL per well. 3. Incubate under hypoxic conditions until day 12. 5 Cardiac Induction Stage 3 (Day 12 to Harvest) 1. Prepare the necessary volume of Stage 3 Induction Medium (volume = number of wells × 1 mL) and place in a 37 ºC water bath for 10–15 min.
Cardiac Tissue Engineering: Methods and Protocols by Milica Radisic, Lauren D. Black III