Cell-derived matrices. Part A

Detalles Bibliográficos
Clasificación:Libro Electrónico
Otros Autores: Caballero, David, Kundu, S. C. (Subhas Chandra), Reis, Rui L.
Formato: Electrónico eBook
Idioma:Inglés
Publicado: Cambridge : Academic Press, 2020.
Colección:Methods in cell biology ; v. 156.
Temas:
Extracellular matrix.
Extracellular Matrix
Substance fondamentale (Histologie)
Extracellular matrix
Acceso en línea:Texto completo
Tabla de Contenidos:
  • Front Cover
  • Cell-Derived Matrices
  • Part A
  • Copyright
  • Contents
  • Contributors
  • Preface
  • Section I: Decellularization and characterization methods
  • Chapter 1: A step-by-step protocol for generating human fibroblast cell-derived completely biological extracellular matrix ...
  • 1. Introduction
  • 2. Materials and methods
  • 2.1. PDMS substrate fabrication
  • 2.2. Collagen grafting on PDMS substrates
  • 2.3. Human dermal fibroblast sheet culture and decellularization procedure
  • 3. Evaluation of hDF derived ECM scaffold
  • 3.1. Immunofluorescence imaging
  • 3.2. Field emission scanning electron micrography
  • 4. Discussion
  • 5. Conclusion
  • Acknowledgments
  • References
  • Chapter 2: Preparation of osteogenic matrices from cultured cells
  • 1. Introduction
  • 2. Osteogenic tissue culture and relevant assays
  • 2.1. Human MSC expansion and osteogenic differentiation
  • 2.1.1. Materials
  • 2.1.2. Procedure
  • 2.2. Assays of OPG secretion by enzyme linked immunosorbent assay (ELISA)
  • 2.2.1. Materials
  • 2.2.2. Procedure
  • 2.3. Assays of ALP activity
  • 2.3.1. Materials
  • 2.3.2. Procedure
  • 2.4. Mineralization assays
  • 2.4.1. Materials
  • 2.4.2. Procedure
  • 2.5. Quantitative real-time RT-PCR (qRT-PCR) assays of biomarker transcription
  • 3. Purification and characterization of OECM
  • 3.1. Purification of matrix
  • 3.1.1. Materials
  • 3.1.2. Procedure
  • 3.2. Assays of protein and calcium
  • 3.2.1. Materials
  • 3.2.2. Procedures
  • 3.3. Morphology of OECM
  • 3.4. Composition of OECM
  • 3.4.1. Materials
  • 3.4.2. Procedure
  • 4. Coating of OECM onto growth surfaces
  • 4.1. Solubilization and coating of OECM onto tissue culture plastic
  • 4.1.1. Materials
  • 4.1.2. Procedure
  • 4.2. In situ coating of OECM by culture and decellularization
  • 4.2.1. Materials
  • 4.2.2. Procedure
  • 5. Discussion, future prospects
  • Acknowledgments
  • References
  • Chapter 3: Decellularized hASCs-derived matrices as biomaterials for 3D in vitro approaches
  • 1. Introduction
  • 2. Materials and methods
  • 2.1. Reagents, equipment and material
  • 2.2. CELL-derived ECM production
  • 2.3. CELL-derived ECM production
  • 3. Evaluation of effectiveness of the decellularization
  • 3.1. Reagents, equipment and material
  • 3.2. DNA quantification
  • 3.3. Visualization of nuclear material
  • 3.4. Qualitative evaluation of collagen
  • 4. Discussion
  • 5. Conclusions
  • Acknowledgments
  • References
  • Chapter 4: Characterization of the elastic properties of extracellular matrix models by atomic force microscopy
  • 1. Introduction
  • 2. Material, equipments and reagents
  • 2.1. Equipment
  • 2.1.1. AFM
  • 2.1.2. Optical microscope
  • 2.1.3. Specific equipment for sample preparation
  • 2.2. Material and reagents
  • 2.2.1. Measurement liquid buffer
  • 2.2.2. AFM cantilever-tip
  • 3. Protocol to measure the mechanical properties of ECM models

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