Jeffrey Morgan and Martin Yarmush bring together in Tissue Engineering Methods and Protocols the first major collection of cutting-edge methods developed and used by leading researchers in the tissue engineering field. These hands-on experts describe easily reproducible protocols for the production and evaluation of polymers, scaffolds, and composites, as well as methods for the isolation, culture, and analysis of cells, including the combination of cells with a variety of materials or devices. Quantitative methods designed to assess cell performance and cell function that will also illuminate our understanding of cellular processes and the cellular response to the chemical and mechanical environment of a functional organ/tissue are also offered.

Preface Contributors Pt. I Materials 1 Preparation of Collagen - Glycosaminoglycan Copolymers for Tissue Regeneration 3 2 Preparation and Use of Tethered Ligands as Biomaterials and Tools for Cell Biology 19 3 Nondestructive Evaluation of Biodegradable Porous Matrices for Tissue Engineering 35 4 Fabrication of Biodegradable Polymer Foams for Cell Transplantation and Tissue Engineering 47 5 Formation of Highly Porous Polymeric Foams with Controlled Release Capability: A Phase-Separation Technique 57 6 Magnetic-Induced Alignment of Collagen Fibrils in Tissue Equivalents 67 7 Preparation and Use of Thermosensitive Polymers 75 8 In Situ Photopolymerized Hydrogels for Vascular and Peritoneal Wound Healing 85 9 Fabrication and Implantation of Gel-Filled Nerve Guidance Channels 101 10 Small Animal Surgical and Histological Procedures for Characterizing the Performance of Tissue-Engineered Bone Grafts 121 11 Preparation and Use of Porous Poly([alpha]-Hydroxyester) Scaffolds for Bone Tissue Engineering 133 Pt. II Cells 12 Quantitative Assessment of Autocrine Cell Loops 143 13 Measurement of Recovery of Function Following Whole Muscle Transfer, Myoblast Transfer, and Gene Therapy 155 14 Preparation of Immortalized Human Chondrocyte Cell Lines 173 Pt. III Clinical Applications 15 Isolation and In Vitro Proliferation of Chondrocytes, Tenocytes, and Ligament Cells 195 16 Culture and Identification of Autologous Human Articular Chondrocytes for Implantation 205 17 Organogenesis of Skeletal Muscle in Tissue Culture 217 18 Hepatocyte Primary Cultures: Currently Used Systems and Their Applications for Studies of Hepatocyte Growth and Differentiation 227 19 Formation and Characterization of Hepatocyte Spheroids 245 20 Isolation and Culture of Human Endothelial Cells 253 21 Isolation and Culture of Microvascular Endothelial Cells 261 22 Initiation, Maintenance, and Quantification of Human Hematopoietic Cell Cultures 271 23 Methods to Isolate, Culture, and Study Osteoblasts 293 24 Cryopreservation of Rat Hepatocytes in a Three-Dimensional Culture Configuration Using a Controlled-Rate Freezing Device 303 Pt. IV Cell Material Composites 25 Microencapsulation of Enzymes, Cells, and Genetically Engineered Microorganisms 315 26 Methods for Microencapsulation with HEMA-MMA 331 27 Micropatterning Cells in Tissue Engineering 349 28 Methods for the Serum-Free Culture of Keratinocytes and Transplantation of Collagen - GAG-Based Skin Substitutes 365 29 Use of Skin Equivalent Technology in a Wound Healing Model 391 30 Preparation and Transplantation of a Composite Graft of Epidermal Keratinocytes on Acellular Dermis 407 31 Development of a Bioartificial Liver Device 423 32 Methods for the Implantation of Liver Cells 433 33 Isolation and Long-Term Maintenance of Adult Rat Hepatocytes in Culture 447 34 Design and Fabrication of a Small Caliper Hybrid Arterial Bioprosthesis 457 35 Methods for the Immunoisolation and Transplantation of Pancreatic Cells 469 36 Methods for the Study of Nerve Cell Migration and Patterning 483 Pt. V Measurement Techniques 37 Estimating Number and Volume of Islets Transplanted Within a Planar Immunobarrier Diffusion Chamber 497 38 Quantitative Measurement of Cell-Cell Adhesion Under Flow Conditions 507 39 Quantitative Measurement of the Biological Response of Cartilage to Mechanical Deformation 521 40 Measuring Receptor-Mediated Cell Adhesion Under Flow: Cell-Free Systems 543 41 In Vitro and In Vivo Quantification of Adhesion Between Leukocytes and Vascular Endothelium 553 42 Quantitative Measurement of Shear-Stress Effects on Endothelial Cells 577 43 Quantitative Modeling of Limitations Caused by Diffusion 595 Index 607