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The Impact of Food Bioactives on Health
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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
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