Basophils and Mast Cells (2^nd Ed., 2nd ed. 2020)
Methods and Protocols
Methods in Molecular Biology Series, Vol. 2163

This second edition provides updated and new chapters to build on and extend the strengths of the first edition. Chapters guide readers through basic biology of basophils, obtaining the cells by purification, culture of stem cells progenitors, peripheral CD34⁺ stem cell-derived mast cells, basophils from CD34⁺ progenitors, diagnostic applications, gene expression patterns in basophils, roles of basophils in different asthma phenotypes, knockout, and disease models. 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.

Authoritative and cutting-edge, Basophils and Mast Cells: Methods and Protocols, Second Edition aims to ensure successful results in the further study of this vital field.

Part I: Introduction and Historical Background

1. Paradigm Shifts in Mast Cell and Basophil Biology and Function: An Emerging View of Immune Regulation in Health and Disease

Ana Olivera and Juan Rivera

Part II: Obtaining the Cells: Purification, Culture, and Counting

2. Purification of Basophils from Peripheral Human Blood

Franco H. Falcone and Bernhard F. Gibbs

3. Mast Cell Purification Protocols

Jasmine Farrington, Elizabeth P Seward, and Peter T Peachell

4. Generation of a Human Allergic Mast Cell Phenotype from CD133⁺ Stem Cells

Hans Jürgen Hoffmann

5. Generation and Culture of Peripheral CD34⁺ Stem Cell-derived Mast Cells (PSCMCs)

6. Modulating the Human Basophil Phenotype During its Development and Maturation: Basophils Derived from in vitro Cultures of CD34+ Progenitor Cells

Donald MacGlashan Jr.

7. Generation of Mast Cells from Murine Stem Cell Progenitors

Emily J. Swindle

8. Integration of the Human Dermal Mast Cell into the Organotypic Co-culture Skin Model

Jonghui Kim, Sven Guhl, Magda Babina, Torsten Zuberbier, and Metin Artuc

9. The Absolute Basophil Count

Elena Borzova

Part III: Basophil/Mast Cell Lines

10. Mast Cell and Basophils Cell Lines: A Compendium

Egle Passante

Daniel Wan, Xiaowei Wang, Ryosuke Nakamura, Marcos J.C. Alcocer, and Franco H. Falcone

12. Use of Humanized Fluorescent Reporter Cell Line RBL NFAT-DsRed for Detection of Allergen-specific IgE in Patient Sera Using Allergen Microarrays

Marina Kalli, Marcos J. C. Alcocer, Andrew J. Blok, and Franco H. Falcone

13. NPY-mRFP Rat Basophil Leukemia (RBL) Reporter: A Novel, Fast Reporter of Basophil/Mast Cell Degranulation

Nafal Barwary, Daniel Wan, and Franco H. Falcone

14. Use of Engineered Nanoparticles (ENPs) for the Study of High-affinity IgE Receptor FcεRI Receptor Engagement and Rat Basophil Leukemia (RBL) Cell degranulation

Alelwani W, Alharbi RA, Wan D, Vllasaliu D, Falcone FH, and Stolnik S

16. Basophil Activation Experiments in Immediate Drug Hypersensitivity: More Than a Diagnostic Aid

Jessy Elst, Vito Sabato, Margo M Hagendorens, Michel van Houdt, Margaretha A. Faber, Chris H Bridts, Didier G Ebo, and Christel Mertens

Rajia Bahri and Silvia Bulfone-Paus

20. Quantity and Quality of Basophil RNA Depend on the RNA Extraction Technique

Victor G. Bechmann, Anette Teo Hansen, Lars K. Poulsen, and Bettina M. Jensen

21. Measuring Histamine and Cytokine Release from Basophils and Mast Cells

Bettina M. Jensen, Ewa A. Bartko, Katrine Baumann, and Per S. Skov

22. Detection of Mast Cells and Basophils by Immunohistochemistry

23. Detection of Basophils and Other Granulocytes in Induced Sputum by Flow Cytometry

Lionel Rohner and Michaela Fux

24. Mast Cell Migration and Chemotaxis Assayed by Microscopy

Monika Bambousková, Zuzana Rubíková, Lubica Dráberová, Pavel Dráber, and Petr Dráber

25. Basophil Stimulation and Signaling Pathway

26. Gene Silencing Approaches in Mast Cells and Primary Human Basophils

Vadim V. Sumbayev and Bernhard F. Gibbs

27. Identification and Immunophenotypic Characterization of Normal and Pathological Mast Cells

José Mário Morgado, Laura Sánchez-Muñoz, Cristina Teodósio, and Luís Escribano

Part VI: Whole Organism: Disease and Knockout Models

28.Mast Cell Mediated Reactions in vivo

29.Identification of Murine Basophils by Flow Cytometry and Histology

Christian Schwartz and David Voehringer

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts