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Methods of adipose tissue biology. Part B / edited by Ormond A. MacDougald, Department of Molecular and Integrative Physiology, Division of Metabolism, Endocrinology and Diabetes, Department of Internal Medicine, School of Medicine, University of Michigan, Ann Arbor, Michigan, USA.

Contributor(s): MacDougald, Ormond AMaterial type: TextTextSeries: Methods in enzymology ; v. 538.Publisher: San Diego, CA : Elsevier, 2014Edition: First editionDescription: 1 online resource (357 pages) : illustrations (some color)Content type: text Media type: computer Carrier type: online resourceISBN: 9780128003367; 0128003367; 0128002808; 9780128002803Subject(s): Adipose tissues | MEDICAL -- Anatomy | SCIENCE -- Life Sciences -- Human Anatomy & Physiology | Adipose tissuesGenre/Form: Electronic books. | Electronic books. | Ebook. Additional physical formats: No titleDDC classification: 611.01827 LOC classification: QP88.15 | .M48ebOnline resources: Click here to access online
Contents:
Front Cover; Methods of Adipose Tissue Biology, Part B; Copyright; Contents; Contributors; Preface; Chapter One: Preparation and Differentiation of Mesenchymal Stem Cells from Ears of Adult Mice; 1. Introduction; 1.1. EMSC characteristics; 2. EMSC Isolation, Culture, Cryopreservation, and Differentiation; 2.1. Materials; 2.2. Solution preparation; 2.3. Methods; 2.3.1. Isolation of cells from external mouse ears; 2.3.2. Culture of EMSC under undifferentiated conditions; 2.3.3. Cryopreservation of EMSC; 2.3.4. EMSC differentiation; 2.3.4.1. Adipogenic differentiation
2.3.4.2. Chondrogenic differentiation2.3.4.3. Osteogenic differentiation; 2.3.4.4. Myogenic differentiation; 3. Conclusions; Acknowledgments; References; Chapter Two: 3-D Adipocyte Differentiation and Peri-adipocyte Collagen Turnover; 1. Introduction; 1.1. Adipose tissue development in vivo; 1.2. 3-D cell biology; 1.3. Matrix metalloproteinase and collagen remodeling; 1.4. ECM remodeling and adipogenesis; 1.5. ECM remodeling during obesity progression; 2. Materials and Methods; 2.1. 3-D adipogenesis in collagen gels; 2.1.1. Type I collagen preparation; 2.1.1.1. Materials; 2.1.1.2. Method
2.1.2. Isolation of primary vascular stromal cells from WAT2.1.2.1. Materials; 2.1.2.2. Methods; 2.1.3. Embedding preadipocytes in type I collagen gels; 2.1.3.1. Materials; 2.1.3.2. Method; 2.1.4. Induction of adipocyte differentiation; 2.1.4.1. Materials; 2.1.4.2. Method; 2.2. Gene expression analysis of 3-D adipocytes; 2.2.1. Harvesting RNA from adipocytes differentiated in 3-D; 2.3. Assessment of insulin signaling of 3-D adipocytes; 2.3.1. Western blot of 3-D samples; 2.4. Imaging of 3-D adipocytes; 2.4.1. Lipid accumulation; 2.4.2. Protein localization; 2.5. Collagen remodeling in vivo
2.5.1. Sirius red staining to assess fibrillar collagen content2.5.1.1. Materials; 2.5.1.2. Method; 2.5.2. Staining of collagen family members in vitro; 2.5.2.1. Materials; 2.5.2.2. Method; 2.5.3. Whole-mount tissue staining; 2.5.3.1. Method; 2.5.4. MMP-dependent cleavage of collagen fibers; 2.5.4.1. Materials; 2.5.4.2. Method; Acknowledgment; References; Chapter Three: Differentiation of White and Brown Adipocytes from Human Pluripotent Stem Cells; 1. Introduction; 2. Human Pluripotent Stem Cells; 2.1. Required materials; 2.2. Cell culture; 2.2.1. Thawing and plating hPSC cells
2.2.2. Passaging and maintaining hPSC cells3. Differentiation into Mesenchymal Progenitor Cells; 3.1. Required materials; 3.2. EB Formation from hPSCs; 3.3. Mesenchymal progenitor cells from EBs; 4. Differentiation into Adipocytes; 4.1. Required materials; 4.2. Inducible lentivirus production; 4.3. White adipocyte differentiation; 4.4. Brown adipocyte differentiation; References; Chapter Four: Optimal Protocol for the Differentiation and Metabolic Analysis of Human Adipose Stromal Cells; 1. Introduction; 2. Isolation of ASCs from AT; 2.1. Materials; 2.2. Procedures for ASC isolation
Summary: This book is a must-have for anyone interested in obesity or the physiology of white or brown adipose tissues. It contains state-of-the-art methods from researchers that are world leaders in this field. Detailed lab protocols range from methods to visualize adipocytes and adipose tissues in humans and experimental models, to convert stem cells into white and brown adipocytes in vitro, to evaluate aspects of adipocyte metabolism, to inducibly knock out genes in adipose tissues, and to evaluate transcriptional control of adipogenesis on a global scale.
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Includes bibliographical references and index.

Online resource; title from PDF title page (ebrary, viewed February 22, 2014).

This book is a must-have for anyone interested in obesity or the physiology of white or brown adipose tissues. It contains state-of-the-art methods from researchers that are world leaders in this field. Detailed lab protocols range from methods to visualize adipocytes and adipose tissues in humans and experimental models, to convert stem cells into white and brown adipocytes in vitro, to evaluate aspects of adipocyte metabolism, to inducibly knock out genes in adipose tissues, and to evaluate transcriptional control of adipogenesis on a global scale.

Front Cover; Methods of Adipose Tissue Biology, Part B; Copyright; Contents; Contributors; Preface; Chapter One: Preparation and Differentiation of Mesenchymal Stem Cells from Ears of Adult Mice; 1. Introduction; 1.1. EMSC characteristics; 2. EMSC Isolation, Culture, Cryopreservation, and Differentiation; 2.1. Materials; 2.2. Solution preparation; 2.3. Methods; 2.3.1. Isolation of cells from external mouse ears; 2.3.2. Culture of EMSC under undifferentiated conditions; 2.3.3. Cryopreservation of EMSC; 2.3.4. EMSC differentiation; 2.3.4.1. Adipogenic differentiation

2.3.4.2. Chondrogenic differentiation2.3.4.3. Osteogenic differentiation; 2.3.4.4. Myogenic differentiation; 3. Conclusions; Acknowledgments; References; Chapter Two: 3-D Adipocyte Differentiation and Peri-adipocyte Collagen Turnover; 1. Introduction; 1.1. Adipose tissue development in vivo; 1.2. 3-D cell biology; 1.3. Matrix metalloproteinase and collagen remodeling; 1.4. ECM remodeling and adipogenesis; 1.5. ECM remodeling during obesity progression; 2. Materials and Methods; 2.1. 3-D adipogenesis in collagen gels; 2.1.1. Type I collagen preparation; 2.1.1.1. Materials; 2.1.1.2. Method

2.1.2. Isolation of primary vascular stromal cells from WAT2.1.2.1. Materials; 2.1.2.2. Methods; 2.1.3. Embedding preadipocytes in type I collagen gels; 2.1.3.1. Materials; 2.1.3.2. Method; 2.1.4. Induction of adipocyte differentiation; 2.1.4.1. Materials; 2.1.4.2. Method; 2.2. Gene expression analysis of 3-D adipocytes; 2.2.1. Harvesting RNA from adipocytes differentiated in 3-D; 2.3. Assessment of insulin signaling of 3-D adipocytes; 2.3.1. Western blot of 3-D samples; 2.4. Imaging of 3-D adipocytes; 2.4.1. Lipid accumulation; 2.4.2. Protein localization; 2.5. Collagen remodeling in vivo

2.5.1. Sirius red staining to assess fibrillar collagen content2.5.1.1. Materials; 2.5.1.2. Method; 2.5.2. Staining of collagen family members in vitro; 2.5.2.1. Materials; 2.5.2.2. Method; 2.5.3. Whole-mount tissue staining; 2.5.3.1. Method; 2.5.4. MMP-dependent cleavage of collagen fibers; 2.5.4.1. Materials; 2.5.4.2. Method; Acknowledgment; References; Chapter Three: Differentiation of White and Brown Adipocytes from Human Pluripotent Stem Cells; 1. Introduction; 2. Human Pluripotent Stem Cells; 2.1. Required materials; 2.2. Cell culture; 2.2.1. Thawing and plating hPSC cells

2.2.2. Passaging and maintaining hPSC cells3. Differentiation into Mesenchymal Progenitor Cells; 3.1. Required materials; 3.2. EB Formation from hPSCs; 3.3. Mesenchymal progenitor cells from EBs; 4. Differentiation into Adipocytes; 4.1. Required materials; 4.2. Inducible lentivirus production; 4.3. White adipocyte differentiation; 4.4. Brown adipocyte differentiation; References; Chapter Four: Optimal Protocol for the Differentiation and Metabolic Analysis of Human Adipose Stromal Cells; 1. Introduction; 2. Isolation of ASCs from AT; 2.1. Materials; 2.2. Procedures for ASC isolation

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