Thermogenic Fat: Methods and Protocols

ISBN-13: 9781493968190 / Angielski / Twarda / 2017 / 214 str.

Jun Wu

Spis treści

Preface…
Table of Contents…
Contributors…

Part I Isolation and Culture Methods

1. Isolation, Primary Culture, and Differentiation of Preadipocytes from Mouse Brown Adipose Tissue
Wei Gao, Xingxing Kong, and Qin Yang

2. Isolation of Mouse Stromal Vascular Cells for Monolayer Culture
Longhua Liu, Louise D. Zheng, Sarah R. Donnelly, Margo P. Emont, Jun Wu, and Zhiyong Cheng

3. Flow Cytometry Assisted Isolation of Adipose Tissue Derived Stem Cells
Umesh D. Wankhade and Sushil G. Rane

4. Flow Cytometric Isolation and Differentiation of Adipogenic Progenitor Cells into Brown and Brite/Beige Adipocytes
Jochen Steinbring, Antonia Graja, Anne-Marie Jank, and Tim J. Schulz

5. Immuno-Magnetic Isolation and Thermogenic Differentiation of White Adipose Tissue Progenitor Cells
Rohollah Babaeikelishomi, Irem Bayindir-Buchhalter, Irina Meln, and Alexandros Vegiopoulos

6. Isolation of Immune Cells from Adipose TissueJonathan R. Brestoff

7. Differentiation and Metabolic Interrogation of Human Adipocytes
Nicki A. Baker, Lindsey A. Muir, Carey N. Lumeng, and Robert W. O’Rourke

8. Protocols for Generation of Immortalized Human Brown and White Preadipocyte Cell Lines
Farnaz Shamsi and Yu-Hua Tseng

Part II Functional Analysis, Bioengineering, and Other Applications

9. Gene Expression and Histological Analysis of Activated Brown Adipocytes in Adipose Tissue
Yun-Hee Lee

10. Genetic Mouse Models: The Powerful Tools to Study Fat Tissues
Xingxing Kong, Kevin W. Williams, and Tiemin Liu 

11. Genetic Manipulation with Viral Vectors to Assess Metabolism and Adipose Tissue Function
Nicolás Gómez-Banoy and James C. Lo

12. Bioenergetic Analyses in Adipose Tissue
Lawrence Kazak

13. Oxygen Consumption Rate and Energy Expenditure in Mice: Indirect Calorimetry
Eun Ran Kim and Qingchun Tong

14. Isolation and Patch-Clamp of Primary Adipocytes
Yanhui Zhang, Dan Tong, Anil Mishra, Litao Xie, and Rajan Sah

15. In Vitro Approaches to Model and Study Communications Between Adipose Tissue and the Liver
Sean O’Connor and Paul Cohen

16. Identification and Quantification of Human Brown Adipose Tissue
Maria Chondronikola, Craig Porter, John O. Ogunbileje, and Labros S. Sidossis

17. Designing 3D Adipospheres for Quantitative Metabolic Study
Takeshi Akama, Brendan M. Leung, Joe Labuz, Shuichi Takayama, and Tae-Hwa Chun

18. Culture and Sampling of Primary Adipose Tissue in Practical Microfluidic Systems
Jessica C. Brooks, Robert L. Judd, and Christopher J. Easley

19. Using Thermogenic Beige Cells to Identify Biologically Active Small Molecules and Peptides
Ling Wu and Bin Xu

Blurb

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. 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.

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.