The Impact of Food Bioactives on Health

Cells Present in the Intestine Role of Microbiota Definition of Concepts: Bioavailability, Bioaccessibility and Bioactivity Static Methods Solubility/Dialyzability Digestion Conditions Applications: Advantages and Disadvantages Static Versus In Vivo Digestion: Conclusions References InfoGest Consensus Method Introduction The Oral Phase The Gastric Phase The Small Intestinal Phase Practicalities Sampling Conclusions References Approaches to Static Digestion Models Introduction Static Models for Protein Hydrolysis Static Models for Lipid Hydrolysis Other Static Models References Geometry Physical Forces Biochemistry References The TNO Gastro-Intestinal Model (TIM)Introduction Concept of TIM TIM-1 Tiny TIM Advanced Gastric Compartment (TIM-agc)The Use of TIM to Study the Bio-accessibility of Nutrients Protein Quality Prediction of Glycemic Response Lipids Conclusions References Dynamic Gastric Model (DGM)Origins and Design of the DGM General Protocol for DGM Experiments Uses of the DGM Food-Based Research Pharmaceutical-Based Research Advantages, Disadvantages and Limitations Availability of the System References Human Gastric Simulator (Riddet Model)Origins of the HGS Model Description Gastric Compartment Gastric Motility Gastric Emptying Gastric Secretions Temperature Control Analysis of HGS Biomechanical Relevance Operating Protocol Preparation of a Food Bolus Gastric Processing Uses of the HGS Role of ACW Activity on Food Digestion Role of Food Material Properties Advantages and Limitations Availability of the System References The DIDGI® System Origins and Design of the DIDGI® System Validation of DIDGI® for the Digestion of Infant Formula Protocol for the In Vitro Dynamic Digestion of Infant Formula Using the DIDGI® System In Vivo Digestion of Infant Formula on Piglets Comparison In Vitro/In Vivo Data Advantages, Disadvantages and Limitations Conclusion and Prospects References Medium Quality pH and CO2 Serum Addition The Cell Culture Hood Cell Culture Terminology (Fig. 1)General Cell Culture Protocols Trypsinisation and Subculturing of Cells Passaging of Cells in Suspension Culture Freezing Cells The Thawing and Recovery of Cells Cell Viability Testing Contamination of Cell Cultures References Measurement of Transepithelial Electrical Resistance (TEER)Basic Protocol Calculating Transepithelial Resistance Verification of Monolayer Integrity by Lucifer Yellow Flux Basic Protocol Summary References Caco-2 Cell Line Origin Features and Mechanisms Stability, Consistency and Reproducibility Relevance to Human In Vivo Situation General Protocols for Caco-2 Cells General Maintenance Protocol for Polarizing Caco-2 Cells in Tissue Culture Inserts Troubleshooting Guide for Transport Experiments Across Caco-2 Monolayers Applications Advantages and Disadvantages Conclusion References HT29 Cell Line Origin Features and Mechanisms Stability, Consistency and Reproducibility Relevance to Human In Vivo Situation General Protocol for HT29-MTX Cells Cell Maintenance Protocol Experimental Protocol for Test Compounds Experimental Read Out Functionality Studies Transport Studies Microorganisms Survival, Adhesion or Invasion Conclusions References The IPEC-J2 Cell Line Origin Special Features/Morphology/Receptors Stability/Consistency/Reproducibility of the System Relevance to Human In Vivo Situation General Protocol Culture Conditions Experimental Readout Sample Preparation Conclusion References Co-cultivation of Caco-2 and HT-29MTX Origin, Features and Mechanisms Stability/Consistency and Reproducibility Relevance to the Human In Vivo Situation General Protocol Assess Viability Experimental Readout Advantages, Disadvantages and Limitations Conclusions References Monocytes and Macrophages Dendritic Cells Human Peripheral Blood Mononuclear Cells T Lymphocytes or T-Cells Origin and Some Features of THP-1 and U937 Cells Stability, Consistency and Reproducibility of the System Relevance to Human In Vivo Situation Other Models with the Same Applicability General Protocol of Culturing THP-1 Cells Differentiation of THP-1 and U937 Monocytes into Macrophages Differentiation of THP-1 and U937 Monocytes into Dendritic Cells Controls to Test Viability and Performance of the Model Critical Notes Read-Out of the System References Peripheral Blood Mononuclear Cells Origin Features and Mechanisms Stability, Consistency and Reproducibility Relevance to Human In Vivo Situation General Protocol Study of Proliferative/Cytotoxic Activity Study of Inflammatory Responses Assess Viability Experimental Read Out Advantages, Disadvantages and Limitations of the System Conclusions References PBMC-Derived T Cells Introduction and Origin Features and Mechanisms Applications of T Cell Cultures General Protocol T Cell Isolation Protocols Indirect Positive Isolation of Human CD4+ T Assess Viability Samples Experimental Readouts References Dendritic Cells Origin Features and Mechanisms DC Subsets General Protocols DC Cell Lines Isolating Primary DCs from Blood CD34+-Derived DCs Monocyte-Derived DCs Asses Viability Experimental Readout Co-stimulation Cytokine Production Other DC Readouts In Vitro Studies on Food Bioactives Using DCs (Table 17.3)Critical Notes References Co-culture Caco-2/Immune Cells Origin, Features and Mechanisms Relevance to Human In Vivo Situation Co-culture Caco-2 and Dendritic Cells Co-culture Caco-2 and B-cells (Raji)Stability, Consistency and Reproducibility General Protocol Co-culture of Caco-2/Human Monocyte Derived DCs (Include Contact Dependent Events)Caco-2/Human Monocyte Derived DCs (Soluble Factors)Caco-2/THP-1 (Soluble Factors)Caco-2/PBMCs (Soluble Factors)Caco-2/B Cells Assess Viability Experimental Readout Advantages, Disadvantages and Limitations Conclusions References STC-1 Cells Origin Features and Mechanisms Stability, Consistency and Reproducibility Relevance to Human In Vivo Situation General Protocol Cell Maintenance Protocol Experimental Protocol for Test Compounds Assess Viability Experimental Read out Conclusions References NCI-H716 Cells Introduction Origin Features and Mechanisms Stability/Consistency/Reproducibility Relevance to the Human L-Cell In Vivo General Protocol Cell Maintenance Protocol Experimental Protocol for Test Compounds Assess Viability Experimental Readout Conclusions References Murine GLUTag Cells Introduction Origin Features and Mechanisms Regulation of GLUTag Cell Secretory Activity by Nutrients Regulation of GLUTag Cell Secretory Activity by Other Signals Relevance to the Human Situation (L-Cell In Vivo)General Protocol Maintenance of the GLUTag Cell Line Performance of Secretion and Expression Assays Assess Viability Experimental Readout of the System Conclusions References Intestinal Rings and Intestinal Segments Everted Sac Ussing Chamber Isolated and Perfused Intestinal Segments -D Culture Systems Intestinal Crypt Organoids as Experimental Models Gastrointestinal Organoids General Protocol Small Intestinal Crypt Isolation Small Intestinal Organoid Culture Small Intestinal Organoid Passage Monitoring Viability General Advantages and Limitations of Intestinal Organoid Systems References Porcine Ex Vivo Intestinal Segment Model The Porcine Ex Vivo Intestinal Segment Model General Protocol Sample Analysis Monitoring Viability Incubation with Food Components and Digested Food Samples Readout of the Porcine Ex Vivo Intestinal Segment Model General Advantages and Disadvantages of the Model References Ussing Chamber The Ussing Chamber General Protocol Monitoring Viability Use of Digested Food Samples Readout of the Ussing Chamber System General Advantages, Disadvantages and Limitations of the Ussing Chamber System References Dynamic Fermentation Models Inoculation of Gut Fermentation Models Host-Gut Microbiota Interaction One Compartment Fermentation Model Description of the VTT One Compartment Fermentation Model History of the Model Special Features Validation of the System Relevance to Human In Vivo Situation Quality in Relation to Other Models with the Same Applicability General Protocol Controls: Positive and Negative Read Out of the System Summary of Advantages, Disadvantages and Limitations of the System and Contingency Plan References The TNO In Vitro Model of the Colon (TIM-2)Description of TIM-2 History of the Model Special Features of the Model Stability and Reproducibility of the System Relevance to Human In Vivo Situation Quality in Relation to Other Models with the Same Applicability General Protocol Controls to Test Stability and Performance of the Model Read-Out of the System and How This Information Can Be Used Advantages, Disadvantages and Limitations of the System References The Simulator of the Human Intestinal Microbial Ecosystem (SHIME®)Description of SHIME®History of the Model Special Features of the Model Stability and Reproducibility of the System Relevance to Human In Vivo Situation Quality in Relation to Other Models with the Same Applicability General Protocol Controls to Test Stability and Performance of the Model Read-Out of the System and How This Information Can Be Used Advantages, Disadvantages and Limitations of the System References The Computer-Controlled Multicompartmental Dynamic Model of the Gastrointestinal System SIMGI Description of the Model History and Special Features of the Model Stability and Reproducibility of the System Relevance to Human In Vivo Situation Quality in Relation to Other Models with the Same Applicability General Protocol Controls to Test Stability and Performance of the Model Read-Out of the System Advantages, Disadvantages and Limitations of the System References