This volume provides methods and approaches to study genetic and environmental regulatory controls on odontogenesis. Chapters guide readers through protocols for isolation and characterization of both epithelial and mesenchymal dental cells, methods on isolation, phenotypic characterization, expansion, differentiation, immunofluorescence, in situ hybridization, immunohistochemistry, imaging protocols, rodent dental fluorosis model, 3D assessment of crown size, dental diseases models, next generation sequencing, genetic and epigenetic studies, genome-wide association studies as well as clinical…mehr
This volume provides methods and approaches to study genetic and environmental regulatory controls on odontogenesis. Chapters guide readers through protocols for isolation and characterization of both epithelial and mesenchymal dental cells, methods on isolation, phenotypic characterization, expansion, differentiation, immunofluorescence, in situ hybridization, immunohistochemistry, imaging protocols, rodent dental fluorosis model, 3D assessment of crown size, dental diseases models, next generation sequencing, genetic and epigenetic studies, genome-wide association studies as well as clinical protocols for measurement of early childhood caries and saliva, and supragingival fluids and biofilm collection and subsequent analyses. 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 ontroubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Odontogenesis: Methods and Protocols aims to guide researchers towards elucidating the secrets and mysteries of a fascinating and unique organ, the tooth.
Micro-dissection and Isolation of Mouse Dental Epithelial Cells of Continuously Growing Mouse Incisors.- Establishment of an Immortalized Mouse Bmp2 Knock--Out Dental Papilla Mesenchymal Cell Line.- Establishment of Stable Cell Lines from Primary Human Dental Pulp Stem Cells.- Isolation of Dental Stem Cell Enriched Populations from Continuously Growing Mouse Incisors.- Application of Cell Lineage Tracing Combined with Immunofluorescence in the Study of Dentogenesis.- Tissue Recombination and Kidney Capsule Transplantation Assays for the Study of Epithelial-mesenchymal Interactions.- Dental Mesenchymal Stem Cells: Dental Pulp Stem Cells, Periodontal Ligament Stem Cells, Apical Papilla Stem Cells, Primary Teeth Stem Cells: Isolation, Characterization and Expansion for Tissue Engineering.- Phenotypic Identification of Dental Pulp Mesenchymal Stem/Stromal Cells Subpopulations with Multiparametric Flow Cytometry.- Dental Pulp Stem Cells: Isolation, Characterization, Expansion and Odontoblast Differentiation for Tissue Engineering.- In vitro Analysis of Intramolecular Signaling Events in PDLSCs using Confocal and TIRF Microscopy.- A Mouse Model to Study Reparative Dentinogenesis.- Multiwalled Carbon Nanotubes for Dental Applications.- Peptide-mediated Biomimetic Regrowth of Human Enamel in situ.- Bioengineering Tooth Bud Constructs using GelMA Hydrogel.- Whole-mount In Situ Hybridization of Mouse Embryos Using DIG-labeled RNA Probes.- In Situ Hybridization on Mouse Paraffin Sections Using DIG-labeled RNA Probes.- Methods for in situ Proteins Visualization in Dental Mineralized Tissues.- In Situ Hibridization in Mineralized Tissues: The Added Value of LNA Probes for RNA Detection.- Immunofluorescence Procedure for Developing Enamel Tissues.- Silver-albumin Tissue Staining Protocol to Visualize Odontogenesis in Whole Embryos.- Isolation of SIBLING Proteins from Bone and Dentin Matrix.- Immunohistochemical Co-localization of Amelogenin and Ameloblastin in Developing Enamel Matrix.- The Expression and Purification of Recombinant Mouse Ameloblastin in e-coli.- Protocols for Studying Formation and Mineralization of Dental Tissues in vivo.- Extraction Protocol for Isolating Dentin Matrix Proteins from Developing Teeth.- Purification of Developing Enamel Matrix Proteins using Preparative SDS PAGE.- Using ImageJ (FIJI) to Analyse and Present X Ray CT Images of Enamel.- Scanning Electron Microscopy (SEM) Methods for Dental Enamel.- Microcomputed Tomography Imaging in Odontogenesis Studies.- Transmission Electron Microscopy (TEM) and Scanning Electron Microscopy (SEM) for the Examination of Dental Hard Tissues.- Rodent Dental Fluorosis Model--Extraction of Enamel Organ from Rat Incisors.- Three Dimensional Assessment of Crown Size and Eruption Space for Developing Third Molars: Data Collection Techniques Based on Cone-beam Computed Tomography (CBCT).- Protocols to Study Dental Caries In Vitro: Microbial Caries Models.- In vitro Caries Models for the Assessment of Novel Restorative Materials.- Protocols to Study Dental Caries In Vitro: pH Cycling Models.- In Vivo Rodent Models for Studying Dental Caries and Pulp Disease.- Protocol GenoDENT: Implementation of a New NGS Panel for Molecular Diagnosis of Genetic Disorders with Orodental Involvement.- Protocols for Genetic and Epigenetic Studies of Rare Diseases Affecting Dental Tissues.- Protocols, Methods, and Tools for Genome-wide Association Studies (GWAS) of Dental Traits.- Measurement of Early Childhood Oral Health for Research Purposes: Dental Caries Experience and Developmental Defects of the Enamel in the Primary Dentition.- The Supragingival Biofilm in Early Childhood Caries: Clinical and Laboratory Protocols and Bioinformatics Pipelines Supporting Oral Metagenomics, Metatranscriptomics and Metabolomics Studies of the Oral Microbiome.- Saliva and Gingival Crevicular Fluid (GCF) Collection forBiomarkers Screening.
Micro-dissection and Isolation of Mouse Dental Epithelial Cells of Continuously Growing Mouse Incisors.- Establishment of an Immortalized Mouse Bmp2 Knock--Out Dental Papilla Mesenchymal Cell Line.- Establishment of Stable Cell Lines from Primary Human Dental Pulp Stem Cells.- Isolation of Dental Stem Cell Enriched Populations from Continuously Growing Mouse Incisors.- Application of Cell Lineage Tracing Combined with Immunofluorescence in the Study of Dentogenesis.- Tissue Recombination and Kidney Capsule Transplantation Assays for the Study of Epithelial-mesenchymal Interactions.- Dental Mesenchymal Stem Cells: Dental Pulp Stem Cells, Periodontal Ligament Stem Cells, Apical Papilla Stem Cells, Primary Teeth Stem Cells: Isolation, Characterization and Expansion for Tissue Engineering.- Phenotypic Identification of Dental Pulp Mesenchymal Stem/Stromal Cells Subpopulations with Multiparametric Flow Cytometry.- Dental Pulp Stem Cells: Isolation, Characterization, Expansion and Odontoblast Differentiation for Tissue Engineering.- In vitro Analysis of Intramolecular Signaling Events in PDLSCs using Confocal and TIRF Microscopy.- A Mouse Model to Study Reparative Dentinogenesis.- Multiwalled Carbon Nanotubes for Dental Applications.- Peptide-mediated Biomimetic Regrowth of Human Enamel in situ.- Bioengineering Tooth Bud Constructs using GelMA Hydrogel.- Whole-mount In Situ Hybridization of Mouse Embryos Using DIG-labeled RNA Probes.- In Situ Hybridization on Mouse Paraffin Sections Using DIG-labeled RNA Probes.- Methods for in situ Proteins Visualization in Dental Mineralized Tissues.- In Situ Hibridization in Mineralized Tissues: The Added Value of LNA Probes for RNA Detection.- Immunofluorescence Procedure for Developing Enamel Tissues.- Silver-albumin Tissue Staining Protocol to Visualize Odontogenesis in Whole Embryos.- Isolation of SIBLING Proteins from Bone and Dentin Matrix.- Immunohistochemical Co-localization of Amelogenin and Ameloblastin in Developing Enamel Matrix.- The Expression and Purification of Recombinant Mouse Ameloblastin in e-coli.- Protocols for Studying Formation and Mineralization of Dental Tissues in vivo.- Extraction Protocol for Isolating Dentin Matrix Proteins from Developing Teeth.- Purification of Developing Enamel Matrix Proteins using Preparative SDS PAGE.- Using ImageJ (FIJI) to Analyse and Present X Ray CT Images of Enamel.- Scanning Electron Microscopy (SEM) Methods for Dental Enamel.- Microcomputed Tomography Imaging in Odontogenesis Studies.- Transmission Electron Microscopy (TEM) and Scanning Electron Microscopy (SEM) for the Examination of Dental Hard Tissues.- Rodent Dental Fluorosis Model--Extraction of Enamel Organ from Rat Incisors.- Three Dimensional Assessment of Crown Size and Eruption Space for Developing Third Molars: Data Collection Techniques Based on Cone-beam Computed Tomography (CBCT).- Protocols to Study Dental Caries In Vitro: Microbial Caries Models.- In vitro Caries Models for the Assessment of Novel Restorative Materials.- Protocols to Study Dental Caries In Vitro: pH Cycling Models.- In Vivo Rodent Models for Studying Dental Caries and Pulp Disease.- Protocol GenoDENT: Implementation of a New NGS Panel for Molecular Diagnosis of Genetic Disorders with Orodental Involvement.- Protocols for Genetic and Epigenetic Studies of Rare Diseases Affecting Dental Tissues.- Protocols, Methods, and Tools for Genome-wide Association Studies (GWAS) of Dental Traits.- Measurement of Early Childhood Oral Health for Research Purposes: Dental Caries Experience and Developmental Defects of the Enamel in the Primary Dentition.- The Supragingival Biofilm in Early Childhood Caries: Clinical and Laboratory Protocols and Bioinformatics Pipelines Supporting Oral Metagenomics, Metatranscriptomics and Metabolomics Studies of the Oral Microbiome.- Saliva and Gingival Crevicular Fluid (GCF) Collection forBiomarkers Screening.
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