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Methods of adipose tissue biology. Part B /
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 a...
| Cote: | Libro Electrónico |
|---|---|
| Autres auteurs: | |
| Format: | Électronique eBook |
| Langue: | Inglés |
| Publié: |
San Diego, CA :
Elsevier,
2014.
|
| Édition: | First edition. |
| Collection: | Methods in enzymology ;
v. 538. |
| Sujets: | |
| Accès en ligne: | Texto completo Texto completo |
Table des matières:
- 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