Since the publication of the previous editions, there has been increased focus on the use of adipose-derived stromal/stem cells (ASC) and stromal vascular fraction (SVF) cells in three-dimensional hydrogel-based scaffolds for the development of microphysiological systems (MPS) serving as in vitro humanized assays and alternatives to in vivo pre-clinical animal models. This third edition volume discusses of the latest technology and advancements in the field of human-derived ASC and SVF. The chapters in this book are organized into four parts. Part One focuses on human ASC's isolation, characterization, and differentiation. Part Two describes the isolation and characterization of ASC and SVF from canine, feline, and murine tissues. Part Three looks at hydrogels, scaffolds, and microphysiological systems, and Part Four talks about the new assays and applications using ASC. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Isolation of Human Adipose-Derived Stem Cells.- Isolation of Huma n Adipose-Derived Stem Cells from Lipoaspirates.- Isolation of Perivascular Mesenchymal Progenitor Cells from Human Adipose Tissue by Flow Cytometry.- Soft Tissue Reconstruction.- Adult Stem Cells Freezing Processes and Cryopreservation Protocols.- Isolation of Murine Adipose-Derived Stromal/Stem Cells for Adipogenic and Osteogenic Differentiation or Flow Cytometry-Based Analysis.- Isolation of Murine Adipose-Derived Stromal/Stem Cells using an Explant Culture Method.- Canine Adult Adipose Tissue-Derived Multipotent Stromal Cell Isolation, Characterization, and Differentiation.- Feline Adult Adipose Tissue-Derived Multipotent Stromal Cell Isolation and Differentiation.- Manufacturing of a Huma Adipose-Derived Hydrogel.- Quality Control Assessment of Human Adipose-Derived Hydrogels.- Analysis of Biomechanical Properties of Adipose-Derived Hydrogelsfor Adipose-Derived Stem Cell Based Hydrogel Culture.- Combination of Adipose Stem Cells and Decellularized Adipose Tissue Hydrogel for Osteogenic Applications.- Preparation of Decellularized Amniotic Membrane and Adipose-Derived Stem Cell Seeding.- Robust Generation of ASC Spheroids for Use as 3D Cultures and in Bioprinted Tissue Models.- Generating and Characterizing Adipose Spheroids from Adipose Stem Cells.- Spheroid Formation of Human Adipose-Derived Stem Cells Using a Liquid-Overlay Technique.- Repurposing Decellularized Lung to Generate Vascularized Fat.- In Vitro Culture of White Adipose Tissue.- Bioreactor Culture to Create Adipose Tissue from Human Mesenchymal Stromal Cells.- A Method for Screening a Kinase Inhibitor Drug Library on ASCs.- Analyses and Utilization of Selectively-Tuned Human Adipose-Derived Stem Cell Exosomes.- Functional Assays in 3D ObaCell® Fat-On-A-Chip Cultures: Lipolysis and Glucose Uptake Assays.- Monitoring ADSC Metabolism Using the Seahorse Analyzer.- Mechanical Stimulation of Adipose-Derived Stem Cells for Functional Tissue Engineering of the Musculoskeletal System via Cyclic Hydrostatic Pressure, Simulated Microgravity, and Cyclic Tensile Strain.- Nuclear Transcription Factor Detection.