Cell Microencapsulation: Methods and Protocols (Methods in Molecular Biology, 1479)

✍ Scribed by Emmanuel C. Opara (editor)

• Publisher: Humana
• Year: 2016
• Tongue: English
• Leaves: 358
• Category: Library

✦ Synopsis

This volume provides a unique forum to review cell microencapsulation in a broad sense by exploring various cell types that have been encapsulated for different purposes, different approaches and devices used for microencapsulation, the biomaterials used in cell microencapsulation, the challenges to the technology, and the current status of its application in different clinical situations. This book is divided in five sections: Section I is an introductory part that discusses historical developments of the technology and its current challenges, as well as the various applications of cell microencapsulation; Section II discusses the main approaches and devices currently used in cell microencapsulation; Section III presents an overview of the various polymeric materials currently in use for cell microencapsulation and the enabling technologies to either monitor or enhance encapsulated cell function; Section IV gives specific examples of the methods used to encapsulate various cell types; and Section V provides an overview of the different clinical situations in which cell microencapsulation has been applied. 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 on troubleshooting and avoiding known pitfalls.

Thorough and practical, Cell Microencapsulation: Methods and Protocols is a valuable reference for researchers, engineers, clinicians, and other healthcare professionals, as well as food technologists who will find detailed descriptions of methods for the microencapsulation of specific cell types and their current of potential clinical and industrial applications. This volume also includes detailed information about the design and manufacture of different devices including large-scale production devices for use in cell microencapsulation.

✦ Table of Contents

Preface
Contents
Contributors
Part I: Introduction
Chapter 1: Historical Perspectives and Current Challenges in Cell Microencapsulation
1 Introduction
2 Macrocapsules and Microcapsules
3 Insufficient Supply of Nutrients is Achilles Heel of Encapsulation
4 Biocompatibility and Biotolerability
5 Impurities Revisit
6 Strategies to Improve Tolerability
7 Biotolerability from the Inside Out
8 Concluding Remarks and Future Considerations
References
Chapter 2: Applications of Cell Microencapsulation
1 Introduction
2 Cell Immunoisolation
3 Immobilization of Cells for Localized Delivery of Therapeutic Products
4 Compartmentalization of Cells in Multilayered Tissue Engineering
5 Expansion of Cells in Culture
6 Cell Microencapsulation in Cancer Treatment
7 Microencapsulation of Bacteria Cells for Probiotic Applications
8 Other Industrial Applications
9 Summary
References
Part II: Approaches to Cell Microencapsulation
Chapter 3: Cell Microencapsulation: Dripping Methods
1 Introduction
1.1 General Equipment
1.2 Protocol
2 Dropwise Regime Under Simple Gravity
2.1 Equipment
2.2 Protocol
3 Dropwise Regime Under Electrostatic Potential: Electro-dripping
3.1 Equipment
3.2 Protocol
4 Dropwise Regime with Coaxial Air Flow
4.1 Equipment
4.2 Protocol
5 Jet-Breaking Regime Using Vibrating Nozzle
5.1 Equipment
5.2 Protocol
6 Jet-Breaking Regime Using a Rotating Wheel
6.1 Equipment
6.2 Protocol
References
Chapter 4: Field Effect Microparticle Generation for Cell Microencapsulation
1 Introduction
1.1 Background
1.2 Electric Field Microparticle Generator
1.3 Factors Influencing Calcium Alginate Beads
1.3.1 Field Strength and Distance
1.3.2 Needle Gauge
1.3.3 Cell Density of Sodium Alginate
1.3.4 Speed of Sodium Alginate Protrusion
1.3.5 Concentration of Alginate Solution
1.3.6 Effect of Field Strength on Viability of NS-1 Cells and Rat Islets
1.3.7 Skin Effect (Gauss’ Law)
1.4 The Advantages of Using an Electric Field Microparticle Generator
2 Materials
2.1 Microenca psulation
2.2 Preparation of NS-1 Cells
2.3 Preparation of Islets
2.4 Diabetic Induction
3 Methods
3.1 Preparation of the Reagents
3.2 Isolation of Rat Islets
3.3 Diabetes Induction and Intraperitoneal Islet Implantation
3.4 Blood Glucose Monitoring
3.5 Measurement of the Diameter of Calcium Alginate Microspheres
3.6 Microenca psulation of Islets
4 Notes
References
Chapter 5: Microfluidic Approach to Cell Microencapsulation
1 Introduction
2 Materials
2.1 Chemicals
2.2 Equipment
3 Methods
4 Notes
References
Part III: Biomaterials and Enabling Technologies in Cell Microencapsulation
Chapter 6: Polymeric Materials for Cell Microencapsulation
1 Introduction
2 Materials
2.1 Tissue Culture Equipment
2.2 Tissue Culture Reagents and Supplies
2.3 Microenca psulation Equipment
2.4 Microencap sulation Reagents and Supplies
2.5 Microencap sulation Working Solutions
2.6 Solutions to Assess Microencapsulated Cell Viability and Encapsulation Efficiency
3 Methods
3.1 Formation of Microspheres or Microcapsules
3.2 Determining Encapsulation Efficiency
3.2.1 MTS Assay
3.2.2 Analysis of Results
3.3 Determining Cell Viability
3.3.1 Live-Dead Assay
3.3.2 Imaging
3.3.3 Analysis
4 Notes
References
Chapter 7: Polymeric Materials for Perm-Selective Coating of Alginate Microbeads
1 Introduction
2 Materials
2.1 Equipment
2.2 Chemicals/Biochemicals
2.3 Solutions
2.4 Animals
3 Methods
3.1 Islet Microencapsulation with the 2-Channel Air-Jacket Device
3.2 Generation of Empty Alginate Microbeads with the Microencapsulator
3.3 Evaluation of Microbead Permeability with Fluorescent Lectins [9]
3.4 Analysis of Changes in Microbead Size [10]
3.5 Implantation of PLL- and PLO-­Coated Microbeads in Normal Rats for the Assessment of Fibrotic Response [11]
3.6 Results from Our Studies
4 Notes
References
Chapter 8: Determination of the Mechanical Strength of Microcapsules
1 Introduction
1.1 Background
1.2 Microcapsule Composition
1.3 Cross-­Linking Cation
1.4 Polyamino Coating
1.5 Microcapsule Morphometry
1.6 Assessment of Microcapsule Mechanical Strength
2 Materials
2.1 Chemicals and Solutions
2.2 Equipment
3 Methods
3.1 Generation of Alginate Microcapsules
3.2 Induction of Mechanical Stress Using Osmotic Pressure
3.3 Induction of Mechanical Stress Using Bead Agitation
4 Notes
References
Chapter 9: The Diffusive Properties of Hydrogel Microcapsules for Cell Encapsulation
1 Introduction
1.1 Hydrogel Encapsulation
1.2 Microcapsule Formulations and Diffusion
1.3 Permselective Hydrogel Membrane Diffusion Testing Models
1.4 Micro-encapsulation of FITC-Dextrans
2 Materials
2.1 Buffers and Solutions
2.2 Kits
2.3 Special Equipment
3 Methods
3.1 Viscometry
3.2 Characterization of Contaminants
3.3 Micro-encapsulation
3.4 Release Studies
3.5 Transmission Electron Microscopy to Evaluate APA Wall Ultrastructure
4 Notes
References
Chapter 10: Methods for Incorporating Oxygen-Generating Biomaterials into Cell Culture and Microcapsule Systems
1 Introduction
2 Materials
2.1 Materials for Oxygen-­Generating Culture System
2.2 Materials for Oxygen-­Generating Microcapsules
3 Methods
3.1 Oxygen-
3.2 Oxygen-
4 Notes
References
Chapter 11: Noninvasive Tracking of Alginate-Microencapsulated Cells
1 Introduction
2 Materials
2.1 Reagents
2.2 Equipment
2.3 Buffer and Solutions
3 Methods
3.1 Solutions
3.2 Electrostatic Droplet Generator Set-Up
3.3 Cell Encapsulation
3.4 Application of Polycation Coat
3.5 Application of Secondary Alginate Layer
3.6 Incorporation of Contrast in Primary Alginate
3.7 Examples of X-ray Contrast, 1H MR Contrast, and 19F MR Contrast
3.8 Adding Cells to Primary Alginate/Contrast Solution
4 Notes
References
Chapter 12: Retrieval of Microencapsulated Islet Grafts for Post-­transplant Evaluation
1 Introduction
2 Materials
2.1 Materials for Islet Microencapsulation
2.2 Materials for Microcapsule Implantation
2.3 Materials for Omentum Retrieval
2.4 Live/Dead Staining Reagents
2.5 Dithizone Staining Reagents
2.6 Dynamic Perifusion Reagents
3 Methods
3.1 Islet Microencapsulation
3.2 Microcapsule Implantation
3.3 Retrieval of Omentum Pouch and Isolation of Microcapsules
3.4 Live/Dead Staining of Retrieved Islets
3.5 Dithizone Staining of Retrieved Islets
3.6 Dynamic Perifusion of Islets and Measurement of Insulin Secretion
4 Notes
References
Part IV: Isolation of Cells for Microencapsulation
Chapter 13: A Method of Porcine Pancreatic Islet Isolation for Microencapsulation
1 Introduction
2 Materials and Methods
2.1 Equipment
2.2 Chemicals/Biochemicals
2.3 Solutions
2.4 Animals
2.5 Methods
3 Results from Our Studies
4 Notes
References
Chapter 14: Selective Osmotic Shock (SOS)-Based Islet Isolation for Microencapsulation
1 Introduction
2 Materials
2.1 Equipment
2.2 Reagents
3 Methods
3.1 Preparation of Reagents
3.2 Isolation of Islets
3.3 Micro-encapsulation of Islets
4 Notes
References
Chapter 15: Preparation and Characterization of Alginate–Chitosan Microcapsule for Hepatocyte Culture
1 Introduction
2 Materials
2.1 Buffers and Solutions
2.2 Kits
2.3 Special Equipment
3 Methods
3.1 Isolation of Porcine Hepatocytes
3.2 Production of Hepatocytes Encapsulation
3.3 Evaluation of Encapsulations and Hepatocytes
3.3.1 Microcapsule Mechanical Stability
3.3.2 Microcapsule Solute Permeability
3.3.3 MTT Assay for Cell Viability [9]
3.3.4 Albumin Assay and Urea Production Assay
3.3.5 Cytochrome P450 1A2 and 3A4 Metabolic Activity Assay
3.3.6 Real-Time PCR
3.3.7 Western Blot
4 Notes
References
Chapter 16: Use of Flow Focusing Technique for Microencapsulation of Myoblasts
1 Introduction
2 Materials
2.1 Myoblast Culture
2.2 Cell Encapsulation
2.3 Microcapsules Coatings
2.4 Viability Assessment of Encapsulated Cells
3 Methods
3.1 Myoblast Culture
3.2 Cell Suspension in Alginate Preparation
3.3 Cell Encapsulation
3.4 Micro-encapsulated Cells Coating
3.5 Viability Assessment of Encapsulated Cells Under Fluorescent Microscope
3.6 Viability Quantification of Encapsulated Cells
4 Notes
References
Chapter 17: Alginate Microbeads for Cell and Protein Delivery
1 Introduction
1.1 Encapsulation of Cells Within the Alginate Beads
1.2 Multilayered Alginate Beads
2 Materials
2.1 Encapsulation of Cells Within Alginate Beads
2.2 Encapsulation of Proteins in Multilayered Alginate Beads
3 Methods
3.1 Encapsulation of Cells Within Alginate Beads (Fig. 1)
3.2 Encapsulation of Proteins in Multilayered Alginate Beads (Fig. 2)
4 Notes
References
Chapter 18: Compartmentalization of Two Cell Types in Multilayered Alginate Microcapsules
1 Introduction
2 Materials
2.1 General Equipments
2.2 Chemicals and Materials
2.3 Cells
2.4 Reagents and Solutions
2.4.1 Cell Culture
2.4.2 Fluorescent Staining
2.4.3 Encapsulation
3 Methods
3.1 Cell Isolation and Purification
3.2 Culture of Granulosa and Theca Cells
3.3 Fluorescent Staining of GC and TC with Cell Tracker Dyes
3.4 Fabrication of Multilayered Alginate Microcapsules
3.5 Imaging the Compartmentalized Cells by Confocal Microscopy
4 Notes
References
Chapter 19: Primary Choroid Plexus Tissue for Use in Cellular Therapy
1 Introduction
2 Materials
2.1 Chemicals
2.2 Special Equipment
3 Methods
3.1 Choroid Plexus Isolation
3.2 Evaluation of Encapsulated Choroid Plexus
3.3 In Vitro Biological Activity of Choroid Plexus-­Conditioned Media
3.4 CP Neurosurgical implantation
3.5 Tissue Preparation of the Lesioned Striatum
4 Notes
References
Chapter 20: Microencapsulation of Stem Cells for Therapy
1 Introduction
2 Materials
2.1 Alginate Microbeads
2.2 Controlled Release Microbeads
3 Methods
3.1 Alginate Microbeads
3.2 Controlled Release Microbeads
3.3 Advantages
3.4 Methods for Analysis
3.4.1 Quantifying Growth Factors Secreted by Microencapsulated Stem Cells
3.4.2 Quantifying mRNA Levels of Micro-encapsulated Cells
3.4.3 Cell Release
3.4.4 Live/Dead Analysis
3.4.5 In Vivo Imaging
4 Notes
References
Chapter 21: Microencapsulated Cells for Cancer Therapy
1 Introduction
2 Materials
2.1 Hybridoma Cell Lines
2.2 Cell Encapsulation
2.3 Microcapsule Coatings
2.4 Assessment of the Viability of Encapsulated Cells
3 Methods
3.1 Hybridoma Cells Culture
3.2 Preparation of the Hybridoma Cell Suspension in Alginate for Microencapsulation
3.3 Hybridoma Cells Microencapsulation
3.4 Alginate-Poly-l-
3.5 Viability Assessment of Encapsulated Cells Under Fluorescent Microscope
3.6 Viability Assessment of Encapsulated Cells in an Automated Cell Counter
3.7 Determination of the Metabolic Activity of the Enclosed Hybridoma Cells
4 Notes
References
Chapter 22: Microencapsulation of Bacterial Cells by Emulsion Technique for Probiotic Application
1 Introduction
2 Materials
2.1 Chemicals and Cultures
2.2 Equipment
3 Methods
3.1 Preparation of Bacterial Cell Suspension
3.2 Preparation of Alginate Solution and Soybean Oil
3.3 Calcium Chloride Solution
3.4 Micro-encapsulation
4 Notes
References
Part V: Cell Microencapsulation in Clinical Applications
Chapter 23: Microencapsulation of Islets for the Treatment of Type 1 Diabetes Mellitus (T1D)
1 Introduction
1.1 Mapping the Field of Interest
1.2 Diabetes Mellitus: A Dangerous Metabolic Disease and a Global Epidemics
1.3 Insulin Therapy Regimens for the Therapy of Diabetes
1.4 New Strategies for the Cure of T1D
1.5 The Concept of Immunoisolation of Islet Cell Grafts
1.6 Microencap
1.7 Alginates for Microencapsulation
1.8 The University of Perugia Microencapsulation Technology for Human Application
2 Materials
2.1 Examination of Isolated Islet Morphology
3 Methods
3.1 Alginic Polymers Qualification for Production of Microcapsules for Human Use
3.2 Islet Cells Quality Control
3.3 In Vitro Glucose-
3.4 Alginate Encapsulation Procedure
3.5 Transplantation Procedures
3.6 Outcome Measures for Assessing Efficacy
3.6.1 Quantitative Measures for Assessment of Efficacy
3.6.2 Outcome Measures for Assessment of Safety
4 Notes
References
Chapter 24: Immunological Challenges Facing Translation of Alginate Encapsulated Porcine Islet Xenotransplantation to Human Clinical Trials
1 Introduction
2 Key Factors Impacting Transplant Outcomes in Alginate Encapsulated Porcine Islet Xenotransplantation
2.1 Alginate Purity
2.2 Polymer Composition and Morphology
3 Antigen Shedding and Danger Associated Molecular Patterns
3.1 Porcine Neonatal Pancreatic Cell Clusters [NPCCs]
3.2 Fetal Porcine Islet-Like Cell Clusters
3.3 Adult Porcine Islets [APIs]
4 Understanding Xenograft Rejection in Nonhuman Primate Recipients
5 Role of Immunomodulation in Preventing Xenograft Rejection
5.1 Immunomod
5.2 Localized Immunosuppression Using Chemother
5.3 The Emerging Role of Biological Agents as Immunomodulators
6 Conclusion
References
Chapter 25: Microencapsulation in Clinical Islet Xenotransplantation
1 Introduction
2 Porcine Islet Xenotransplantation
2.1 Initial Trial in Sweden
2.2 Chinese Trial
2.3 Mexican Trial
3 Microencapsulation in Clinical Islet Xenotransplantation (New Zealand, Russian, and Argentinian Trials)
3.1 Initial Case
3.2 Collaboration Between Living Cell Technologies Ltd. (LCT) and the University of Perugia
3.3 Clinical Trials of APA Encapsulated Porcine Islet Xenotransplantation
4 Microencapsulation in Clinical Islet Allotransplantation
4.1 First Microencapsulated Allogeneic Islet Transplantation in the USA
4.2 Australian Trial
4.3 Italian Trial
5 Research Targets for Microencapsulated Porcine Islet Xenotransplantation
5.1 Lessons from the Clinical Trials
5.2 Improving the Quality of Porcine Islets
5.3 Improving Capsules
5.4 Implant Sites
6 Conclusions
References
Chapter 26: Methods for Microencapsulated Porcine Islet Production
1 Introduction
2 Materials
2.1 Porcine Pancreas Procurement and Preservation
2.2 Porcine Islet Isolation and Evaluation: NCGM Method
2.3 Microencap
3 Methods
3.1 Porcine Pancreas Procurement and Preservation
3.2 Porcine Islet Isolation and Evaluation: NCGM Method
3.2.1 Evaluation
3.3 Microencap
4 Notes
4.1 Porcine Pancreas Procurement
4.2 Porcine Islet Isolation
4.3 Microenca
References
Chapter 27: Microencapsulation of Parathyroid Cells for the Treatment of Hypoparathyroidism
1 Introduction
2 Materials
2.1 Cell Culture
2.2 Tissue Cryopreservation
2.3 Encapsulation
3 Methods
3.1 Primary Culture: Cell Suspension
3.2 Cryopreservation
3.2.1 Tissue Cryopreservation Procedure
3.2.2 Cell Cryopreservation Procedure
3.3 Encapsulation Technique
3.3.1 Cell Encapsulation
3.3.2 Tissue Encapsulation
3.4 Transplant
4 Notes
References
Index