Main description:

This volume presents a broad collection of state-of-the-art methods to study the biology of thermogenic fat using in vitro cell culture and animal models. Chapters guide the readers through protocols on differentiation of human pluripotent stem cells and murine adipocyte precursors; methods for measuring mitochondrial respiration, heat generation, brown fat activation, and effects on energy metabolism in mice; and techniques for AAV-mediated gene delivery, transplantation of adipose tissue, isolation of adipose tissue immune cells and extracellular vesicles, and mass spectrometry-based profiling of brown fat lipids. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and methods, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.

Authoritative and cutting-edge, Thermogenic Fat: Methods and Protocols aims to be comprehensive guide for researchers in the field.

Contents:

1 Differentiation of Human Pluripotent Stem cells (hPSCs) into Brown-Like Adipocytes

Stefania Carobbio and Antonio Vidal-Puig

2 Culture and Differentiation of Primary Preadipocytes from Mouse Subcutaneous

White Adipose Tissue

Julia H. DeLuca and Shannon M. Reilly

3 Isolation and Differentiation of Adipocyte Precursors Derived from Neonatal Murine Brown

Adipose Tissue

Yali Chen and Anyuan He

4 Determination of Adipocyte Size and Number

Terri A. Pietka and Nada A. Abumrad

5 Measuring Mitochondrial Uncoupling in Mouse Primary Brown Adipocytes Differentiated ex vivo

Michaela Veliova, Marcus J. Tol, Peter Tontonoz, Orian S. Shirihai, and Marc Liesa

6 Assessment of Bioenergetics in BAT Mitochondria

Alek D. Peterlin, Jordan M. Johnson, and Katsuhiko Funai

7 Use of Isothermal Microcalorimetry to Measure Cellular Heat Production in Thermogenic Adipocytes

Muhammad Hamza Bokhari Carina Halleskog, Irina Shabalina, and Tore Bengtsson

8 Measurement of Intracellular Temperature in Brown Adipocytes using a Cationic

Fluorescent Polymeric Thermometer

Toshikazu Tsuji, Kazuaki Kajimoto, and Noriko Inada

9 Indirect Calorimetry to Assess Energy Balance in Mice: Measurement and Data Analysis

William Bernard Rubio, Marissa D. Cortopassi, and Alexander S. Banks

10 Thermogenic Phenotyping in Mice

Donghua Hu and Irfan J. Lodhi

11 Calculating Diet-Induced Thermogenesis in Mice

Tomomi Yamazaki

12 Assessment of Brown and Beige Adipose Tissue Activation in Mice using PET/CT Imaging

Yu-Jen Chen, Pei-Xin Liao, Wen-Hung Kuo, Ching-Yi Chen, Shih-Torng Ding, and Mei-Hui Wang

13 Imaging Brown Adipose Tissue with TSPO PET Tracers in Preclinical Animal Studies

Biyue Zhu, Steven H. Liang, and Chongzhao Ran

14 Use of CIDEA Reporter Mouse Model for Screening Thermogenic Fat Activating Drugs

Yeonho Son, Cheoljun Choi, and Yun-Hee Lee

15 AAV-Mediated Gene Delivery to Mouse Brown Adipose Tissue

Wei Huang, Rhiannon Bates, and Lei Cao

16 Subcutaneous Transplantation of White Adipose Tissue

Yu-Sheng Yeh, Mari Iwase, Satoko Kawarasaki, Jungin Kwon, Astrid Rodriguez-Velez, Xiangyu Zhang,

Se-Jin Jeong, Tsuyoshi Goto, and Babak Razani

17 Brown Adipose Tissue Transplantation

Subhadra Gunawardana

18 Isolation and Characterization of Brown Adipose Tissue T Cells

Ling-Ran Kong and Cheng-Chao Ruan

19 Isolation of Adipose Tissue Extracellular Vesicles

Jessica M. Sabio and Clair Crewe

20 Isolation and Mass Spectrometry-Based Profiling of Major Lipids in Brown Adipose Tissue

Dongliang Lu, Hideji Fujiwara, Irfan J. Lodhi, and Fong-Fu Hsu

21 Methods for Single Cell Transcriptomic Analysis of Adipose Tissue

Farnaz Shamsi