Download e-book for iPad: Cardiac Tissue Engineering: Methods and Protocols by Milica Radisic, Lauren D. Black III

By Milica Radisic, Lauren D. Black III

ISBN-10: 1493910469

ISBN-13: 9781493910465

ISBN-10: 1493910477

ISBN-13: 9781493910472

Cardiac Tissue Engineering: tools and Protocols presents a set of protocols on cardiac tissue engineering from pioneering and major researchers around the world. those contain equipment and protocols for mobile instruction, biomaterial training, telephone seeding, and cultivation in numerous structures. Written within the hugely profitable Methods in Molecular Biology sequence structure, chapters contain introductions to their respective issues, lists of the required fabrics and reagents, step by step, conveniently reproducible laboratory protocols, and key pointers on troubleshooting and keeping off identified pitfalls.

Authoritative and useful, Cardiac Tissue Engineering: equipment and Protocols highlights the key ideas, either experimental and computational, for the examine of cardiovascular tissue engineering.

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Extra info for Cardiac Tissue Engineering: Methods and Protocols

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Enzymatic dissociation with TrypLE). 2. TrypLE Select. 3. 2, item 8). 4. Hemocytometer. 5. Trypan Blue. 6. Aggrewell™ 400 plates (StemCell Technologies 27845). 7. Aggrewell™ Rinsing Solution (StemCell Technologies 07010). 8. 5 ng/mL BMP4, 10 μM Y-27632 ROCK Inhibitor. 1 mL of medium is required per well of a 24 well Aggrewell™ 400 plate. 9. OPTIONAL: 37 μm strainer. 4 Cardiac Induction Components 1. Stage 1 Induction Medium: Modified StemPro-34, 10 ng/mL BMP4, 5 ng/mL bFGF, 6 ng/mL Activin A.

7. Fill the wells with 150 μl EBM medium with supplements and incubate at 37 °C overnight (should be 18 h, and no more than a 24 h incubation) (see Note 17). 8. Take photographs of the capillary-like structure formations using fluorescence microscopy. Evaluate the number of CACs (blue DAPI+ cells) that have incorporated into the tubule structures (orange HUVECs), as well as the total length of tubule formation (see Note 18) (Fig. 4). 6 DiI and Lectin Staining 1. Prepare (DiI)-labeled acetylated low-density lipoprotein (acLDL) stain (stock at 1 mg/ml).

Transfer the samples to 5 mL round-bottom flow cytometer analysis tubes and analyze on a flow cytometer using the red laser. 4 Notes 1. 3 mL aliquots of the supplement and store at −20 °C. When preparing complete medium from the aliquots, completely thaw 1 aliquot of supplement in a 37 °C water bath and then add the supplement to a 50 mL aliquot of basal medium. 2. To prepare feeder-depleted hESCs, passage the cells onto Growth Factor Reduced Matrigel (BD 354230) coated 6 well plates at a split ratio that will produce 1–2 × 106 cells per well after 1–2 days.

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Cardiac Tissue Engineering: Methods and Protocols by Milica Radisic, Lauren D. Black III


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