Cell Separation

Methods and Selected Applications

1st Edition - December 28, 1982
Editors: Thomas G. Pretlow, Theresa P. Pretlow
Language: English
eBook ISBN:
9 7 8 - 1 - 4 8 3 2 - 1 9 3 8 - 7

Cell Separation: Methods and Selected Applications is a compendium of articles on the design and/or application of methods for the separation of cells. This volume presents… Read more

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Cell Separation: Methods and Selected Applications is a compendium of articles on the design and/or application of methods for the separation of cells. This volume presents contributions on relatively finite subjects on cell separation. It covers topics on cell separation such as methods for obtaining cells in suspension from animal tissues; some of the kinds of data that are helpful in the description of cell purifications; and separation of host cells infiltrating tumors and allografts by velocity sedimentation at unit gravity. The separation of different kinds of nucleated cells from blood by centrifugal elutriation; a new approach to the separation of cells at unit gravity; and the isolation and culture of homogeneous populations of glomerular cell types are elucidated as well. Experimental oncologists, hematologists, immunologists, cell biologists, endocrinologists, and others who are not already expert in the use of methods for cell separation will find the book highly useful.

Contributors

Preface

1. Methods for Obtaining Cells in Suspension from Animal Tissues

I. Introduction

II. Nonenzymatic Methods of Tissue Disaggregation

III. Proteolytic Enzymes for Disaggregation

IV. Subsidiary Enzymes

V. Protease Inhibitors

VI. Protective Agents

VII. Concluding Remarks

References

2. Evaluation of Data, Problems, and General Approach

I. Introduction

II. Markers

III. Quantitative Characterization of Cells Before and After Cell Separation

IV. Morphological Criteria

V. Concluding Comments

References

3. Sedimentation of Cells: An Overview and Discussion of Artifacts

I. Introduction

II. Theory

III. Velocity Sedimentation

IV. Isopycnic Sedimentation

V. Comparison of Velocity and Isopycnic Sedimentation

VI. Differential Sedimentation and Discontinuous Gradients

VII. Artifacts in Sedimentation of Cells

VIII. Concluding Remarks

References

4 . Separation of Host Cells Infiltrating Tumors and Allografts by Velocity Sedimentation at Unit Gravity

I. Introduction

II. Theoretical Considerations

III. Potential Problems and Practical Limitations

IV. Velocity Sedimentation of Dissociated Animal Tumors

v. Velocity Sedimentation of Ascites Tumors, Leukemias, and Lymphomas

VI. Velocity Sedimentation of Disaggregated Human Neoplasms

VII. Recovery of Allograft-infiltrating Host Cells by 1 g Velocity Sedimentation

VIII. Concluding Remarks

References

5. Analytical Characterization of Adult Granulocyte-Macrophage Progenitor Cells by Sedimentation Velocity and Buoyant Density

I. Introduction

II. Purification Studies

III. Characterization of the Clonable Mouse Granulocyte-Macrophage Progenitor Cells (CFU-C)

IV. Relationship of Mouse CFU-C to Other Clonable Hemopoietic Precursors and Stern Cells

V. Physical Properties of Human CIonable Granulocyte and Macrophage Progenitor Cells

VI. Conclusions

References

6. Sedimentation of Cells in Colloidal Silica (Percoll)

I. Introduction

II. Properties of Percoll

III. Principles of Cell Separation in Percoll

IV. Practical Aspects of the Use of Percoll

v. Does Percoll Interfere with Cell Functions?

VI. Future Use of Percoll

References

7. Separation of Different Kinds of Nucleated Cells from Blood by Centrifugal Elutriation

I. Introduction

II. Physical Characteristics of Human Blood Leukocytes

III. Practical Technique

IV. Isolation of Lymphocytes

V. Isolation of Monocytes

VI. Isolation of Granulocytes

VII. Concluding Remarks

References

8. A New Approach to the Separation of Cells at Unit Gravity

I. Introduction

II. Theory

III. CelSep Apparatus for Unit Gravity Separations

IV. Standard Separation Procedure

V. Separation of Human Monocytes from Blood

VI. Characterization of Human Myeloid Stem Cells

VII. Fractionation of Canine Gastric Cells

VIII. Advantages and Disadvantages of the CelSep Method

References

9. Electronic Cell Sorting of Hemopoietic Progenitor Cells

I. Introduction

II. General Considerations

III. Sorting of Unstained Hemopoietic Cells

IV. Sorting of Hemopoietic Cells Labeled with Antibodies

V. Sorting of Hemopoietic Cells Labeled with Lectins

VI. Sorting of Hemopoietic Cells by Other Techniques

VII. Summary and Future Perspectives

References

10. Separation of Individual Cells from the Fundic Gastric Mucosa

I. Introduction

II. Methods for Gastric Mucosal Cell Isolation

III. Methods for Cell Purification

IV. General Characteristics of Isolated Gastric Cells

V. Specific Advantages and Disadvantages of the Various Purification Methods

VI. Functional Characterization of Isolated Gastric Cells

References

11. Isolation and Culture of Homogeneous Populations of Glomerular Cell Types

I. Introduction

II. Background

III. Techniques for Isolating Glomerular Cells

IV. Tissue Culture Medium Used to Culture Glomerular Cells

V. Properties of Glomerular Cells in Culture

VI. Future Areas of Research on the Biology and Pathobiology of Glomerular Cells

References

12. Separation and Subfractionation of Blood Cell Populations Based on Their Surface Properties by Partitioning in Two-Polymer Aqueous Phase Systems

I. Introduction

II. Separation and Subfractionation of Blood Cells

III. Cell-Cell Affinity Reflected by Countercurrent Distribution

IV. Conclusion

References

13. Purification of Basophilic Leukocytes from Guinea Pig and Human Blood and from Guinea Pig Bone Marrow

I. Introduction

II. Purification of Guinea Pig Blood and Bone Marrow Basophils

III. Purification of Human Peripheral Blood Basophils

References

Index