Cell Separation

Methods and Selected Applications

1st Edition - September 28, 1983
Editors: Thomas G. Pretlow, Theresa P. Pretlow
Language: English
eBook ISBN:
9 7 8 - 1 - 4 8 3 2 - 1 9 3 9 - 4

Cell Separation: Methods and Selected Applications, Volume 2 provides information pertinent to the design and application of methods for the separation of cells. This book covers a… Read more

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Cell Separation: Methods and Selected Applications, Volume 2 provides information pertinent to the design and application of methods for the separation of cells. This book covers a variety of topics, including thyroid structure and function, procurement of eosinophils, elutriation, stromal cells, isolation of epithelial cells, and separation of cells. Organized into 15 chapters, this volume begins with an overview of the structure of thyroid parenchyma and the properties of endocrine parafollicular cells. This text then summarizes the various aspects of the separation methodology to assist investigators with the use of elutriation. Other chapters describe the methods that are useful for the separation of stromal cells from tumors. This book discusses as well the types of methods that are available for the purification of cells from tumors. The final chapter deals with the biological significance of the erythroblastic nest. This book is a valuable resource for cell biologists, experimental oncologists, hematologists, immunologists, and endocrinologists.

Contributors

Contents of Volume 1

Preface

1. Separation of Parafollicular Cells from Thyroid Follicular Cells by Affinity Chromatography Using Thyroglobulin-Coupled Sepharose

I. Thyroid Structure and Function

II. Parafollicular Cells and Serotonin

III. Obtaining Parafollicular Cells for Study

IV. Serotonin Binding Protein

V. Distribution of Parafollicular Cells

VI. Dissociation of the Cells of the Thyroid

VII. Separation of Parafollicular Cells

References

2. Procurement and Purification of Eosinophils

I. Introduction

II. Procurement of Eosinophils

III. Purification

IV. Summary and Conclusions

References

3. Experimental Factors Involved in Separation by Centrifugal Elutriation

I. Introduction

II. Methodology of Elutriation

III. Concluding Remarks

References

4. Analysis and Separation of Stromal Cells Infiltrating Tumors

I. Introduction

II. Previous Work

III. Quantitation and Subcompartments

IV. Identification of Stromal Cells and Morphometry

V. Purification of Cells from Tumors

VI. Concluding Comments

References

5. Selective Isolation of Epithelial Cells in Primary Expiant Cultures of Human and Animal Tissues

I. Introduction

II. Initiation of Primary Expiant Cultures

III. Selective Isolation of Epithelial Cells

IV. Identification of Human Bronchial and Bladder Epithelial Cells in Primary Expiant Cultures

V. Discussion

VI. Other Techniques

References

6. Isolation of Human Natural Killer Cells (Large Granular Lymphocytes)

I. Introduction

II. Isolation of Large Granular Lymphocytes (LGL)

III. Comments

References

7. Purification of Islets and Cells from Islets

I. Introduction

II. Isolation of Pancreatic Islets

III. Islet Purification

IV. Preparation of Islet Cells

V. Separation of Islet Cell Subpopulations

VI. Summary

References

8. Magnetic Cell Sorting

I. Introduction

II. Magnetism and Magnetic Forces

III. High Gradient Magnetic Separation

IV. Magnetic Particles for Cell Labeling

V. Conclusions

References

9. Separation of Cells by Preparative Density Gradient Electrophoresis

I. Introduction

II. Electrophoresis Theory

III. Preparative Cell Electrophoresis in Density Gradients

IV. Separation and Characterization of Cells by Density Gradient Electrophoresis

V. Concluding Remarks

References

10. Isolation, Characterization, and Culture of Human Mammary Epithelial Cells

I. Introduction

II. Isolation of Mammary Epithelial Cells in Culture

III. Method for the Culture of Human Mammary Epithelial Cells

IV. Verification of the Mammary Epithelial Origin of the Cultured Cells

V. Verification of the Malignant Origin of the Epithelial Cells Derived from Tumor Specimens

VI. A Marker of Early Stages in Malignant Progression

VII. Use of the Clonal Assay to Study Chemotherapeutic Drug Sensitivity

VIII. Summary

References

11. Solid-Phase Fractionation of Lymphoid Cells on Ligand-Coated Plastic Plates

I. Introduction

II. Lymphocyte Fractionation

III. Isolation of Surface Immunoglobulin-Positive Lymphocytes

IV. Isolation of Human Fc Receptor-Bearing Lymphocytes on Antigen-Antibody Coated Dishes

V. Isolation of Fc Receptor-Bearing Human T Lymphocytes

VI. Isolation of Antigen-Specific Lymphocytes

VII. Isolation of Lymphocyte Subsets with Monoclonal Antibodies

VIII. Summary

References

12. Large Capacity Separation of Cells in a Reorienting Zonal Rotor

I. Introduction

II. General Characteristics of Reorienting Zonal Rotors

III. Types of Separations Carried out in the TX-28 Rotor

IV. Choice of Reorienting Zonal Rotor Versus Other Techniques

V. Concluding Comments

References

13. Short-Term Culture of Cells from Human Solid Tumors in Semisolid Medium

I. Introduction

II. Methodology for Clonogenic Assays

III. Conclusions

References

14. Preparation and Performance of the Erythrocyte-Unsaturated Fatty Acid (E-UFA) Test

I. Historical Introduction

II. Technique of Red Blood Cell (RBC) Preparation

III. Measurement of Absolute Mobility

IV. Precautions: Interfering Effects of Medication

V. Some Unexpected Results

VI. "Silent" and Subclinical Multiple Sclerosis

VII. Etiology and Pathogenesis of Multiple Sclerosis

VIII. Summary

References

15. Isolation of Erythroblastic Nests and Separation of Their Cellular Components

I. Introduction

II. Definitions

III. Why Erythroblastic Nest (EN)?

IV. Method and Guidelines

V. Generation of Erythropoietic Spleens Containing ENs

VI. Preparation of Cell Suspensions from Erythropoietic Spleens

VII. Preparation of Spleen Cell Suspensions Enriched in ENs

VIII. EN Purification

IX. EN Dissociation

X. Separation of EN Central Cells from Erythroblasts

XI. Summation of Methods and Selected Applications

XII. Conclusions

XIII. Perspectives for the Future

References

Index