Read more
This volume presents a collection of protocols that describe methodologies to study thermogenic fat biology from various angles. This book is divided into 2 parts. Part 1 focuses on establishing in vitro culture systems. The chapters in this section introduce techniques on how to isolate, culture, and differentiate primary fat cells from both laboratory mice and humans. This part also presents flow cytometry methods to isolate various subpopulations of precursors within the stromal vascular fraction of the adipose tissue, which contains both preadipocytes and immune cells. Part 2 introduces multiple means to genetically manipulate and evaluate brown and beige fat in vivo. The chapters in this section explore methods on bioenergetics analyses both in vitro and in vivo. They also cover how to evaluate thermogenic fat contents and activity in humans, how to culture these cells though interdisciplinary approaches, and how to use thermogenic fat cell lines to carry out drug screens. Writtenin 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.
Thorough and cutting-edge, Thermogenic Fat: Methods and Protocols is a valuable resource for both experts and novices who are interested in learning assays and investigating brown and beige fat functions.
List of contents
Isolation, Primary Culture, and Differentiation of Preadipocytes from Mouse Brown Adipose Tissue.- Isolation of Mouse Stromal Vascular Cells for Monolayer Culture.- Flow Cytometry Assisted Isolation of Adipose Tissue Derived Stem Cells.- Flow Cytometric Isolation and Differentiation of Adipogenic Progenitor Cells into Brown and Brite/Beige Adipocytes.- Immuno-Magnetic Isolation and Thermogenic Differentiation of White Adipose Tissue Progenitor Cells.- Isolation of Immune Cells from Adipose Tissue.- Differentiation and Metabolic Interrogation of Human Adipocytes.- Protocols for Generation of Immortalized Human Brown and White Preadipocyte Cell Lines.- Gene Expression and Histological Analysis of Activated Brown Adipocytes in Adipose Tissue.- Genetic Mouse Models: The Powerful Tools to Study Fat Tissues.- Genetic Manipulation with Viral Vectors to Assess Metabolism and Adipose Tissue Function.- Bioenergetic Analyses in Adipose Tissue.- Oxygen Consumption Rate and Energy Expenditure in Mice: Indirect Calorimetry.- Isolation and Patch-Clamp of Primary Adipocytes.- In Vitro Approaches to Model and Study Communications Between Adipose Tissue and the Liver.- Identification and Quantification of Human Brown Adipose Tissue.- Designing 3D Adipospheres for Quantitative Metabolic Study.- Culture and Sampling of Primary Adipose Tissue in Practical Microfluidic Systems.- Using Thermogenic Beige Cells to Identify Biologically Active Small Molecules and Peptides.
Summary
This volume presents a collection of protocols that describe methodologies to study thermogenic fat biology from various angles. This book is divided into 2 parts. Part 1 focuses on establishing in vitro culture systems. The chapters in this section introduce techniques on how to isolate, culture, and differentiate primary fat cells from both laboratory mice and humans. This part also presents flow cytometry methods to isolate various subpopulations of precursors within the stromal vascular fraction of the adipose tissue, which contains both preadipocytes and immune cells. Part 2 introduces multiple means to genetically manipulate and evaluate brown and beige fat in vivo. The chapters in this section explore methods on bioenergetics analyses both in vitro and in vivo. They also cover how to evaluate thermogenic fat contents and activity in humans, how to culture these cells though interdisciplinary approaches, and how to use thermogenic fat cell lines to carry out drug screens. Writtenin 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.
Thorough and cutting-edge, Thermogenic Fat: Methods and Protocols is a valuable resource for both experts and novices who are interested in learning assays and investigating brown and beige fat functions.
