Methods in Cell Biology: Animal Cell Culture Methods

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ISBN: 9780125441599 | 0125441592
Cover: Hardcover
Copyright: 7/1/1998

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This volume provides complete and thorough coverage of the classical and state-of-the-art methods used in cell culture. It also includes basic principles used in the selection of cells for specific scientific study, as well as analytical and procedural techniques. Key Features * Reviews basic principles of cell culture * Gives options and techniques on how to look at cells

Contributors

(2)

Preface

xiii

SECTION I Principles of Cell Culture

(67)

1. Animal Cell Culture Equipment and Techniques

(17)

Angela Helmrich

David Barnes

I. Introduction

(1)

II. Equipment

(6)

III. Laboratory Design

(1)

IV. Materials

(2)

V. Cell Culture Methods

(4)

References

(3)

2. Making Informed choices: Medium, Serum, and Serum-Free Medium How to Choose the Appropriate Medium and Culture system for the Model You Wish to Create

(11)

Jennie P. Mather

I. Introduction

(1)

II. The Role of Medium

(2)

III. pH Control

(1)

IV. Selecting the Appropriate Medium

(1)

V. Screening Conditioned Medium for Biological Activity

(1)

VI. Media Preparation

(1)

VII. Serum, Plasma, and Other Undefined Additives

(1)

VIII. Testing Media and Components and Quality Control: "It's in the Water"

(1)

IX. Troublesbooting Medium Problems

(1)

X. Altering Commercial Media for Special Uses

(1)

XI. Medium Optimization

(1)

XII. Choosing the Optimal Medium: The "Quick and Dirty" Method

(1)

References

(2)

3. Cell Line Availability: Where to Get the Cell Lines You Need

(19)

Robert J. Hay

I. Introduction

(1)

II. Diversity and Selection

(4)

III. Nomenchature

(1)

IV. Commaercial Utility

(1)

V. AExample Cell Lines Developed

(2)

VI. Cell Line Cryopreservation and Authentication

(6)

References

(4)

4. Cell Culture Contamination: Sources, Consequences, Prevention, and Elimination

(20)

Carolyn Kay Lincoln

Michael G. Gabridge

I. Introdiction

(1)

II. Cell Culture Technique

(2)

III. Cell Culture Contamination

(11)

IV. Summary

(1)

References

(6)

SECTION II Establishing Cell Lines

(150)

5. Immoratalization by Gene Transfection

(24)

Yoshinori Katakura

Shahabuddin Alam

Sanetaka Shirahata

I. Genes Used for Cell Immortalization

(7)

II. Gene Delivery

(2)

III. Practical Procedures for Human T-Cell Immoratalization by Oncogene Transfection

(4)

IV. Conclusions

(2)

References

(9)

6. Establishment of Mammalian Testicular Cell Lines

(18)

Marie-Claude C. Hoffmann

Jose Luis Millan

I. Introduction

(1)

II. Establishment of Germ Cell and Testicilar Cell Lines Using SV40 Large T Antigen

(6)

III. In Vitro Seminiferous Cords and Their Use for Promoter Analysis of Germ Cell-Specific Genes

101

(4)

IV. Establishment of Germ Cell Lines Using a Combination of the SV40 Large T Antigen and Temperature-Sensitive p53

105

(2)

V. Conclutions

107

(2)

References

109

(3)

7. Cell Hybridization, Hybridomas, and Human Hybridomas

112

(35)

Sanetaka Shirahata

Yoshinori Katakura

Kiichiro Teruya

I. Introdiction

112

(2)

II. Cell Hybridization

114

(4)

III. Hybridomas

118

(9)

IV. Human Hybridomas

127

(13)

V. Application of Human Monoclonal Antibodies

140

(2)

References

142

(5)

8. Establishing Human Glioma-Derived Cell Lines

147

(21)

Manfred Westphal

Hildegard Meissner

I. Introdiction

147

(1)

II. Culture Methodology and General Aspects

148

(3)

III. Cell Culture Methodology, Results, and Application

151

(8)

IV. Discussion, Review, and Perspectives

159

(4)

References

163

(5)

9. Culture Methods for selective Growth of Normal Rat and Human Schwann Cells

168

(20)

Ronghao Li

I. Introduction

168

(2)

II. Meterials and Instruments

170

(3)

III. Culture of Rat Schwann Cells from Dorsal Root Ganglia (DRG)

173

(6)

IV. Culture of Adult human Schwann Cells

179

(2)

V. Characteristics of Schwann Cell Cultures

181

(4)

References

185

(3)

10. Invertebrate Cell Culture Considerations: Insects, Ticks, Shellfish, and Worms

188

(16)

Christopher J. Bayne

I. Introduction

188

(2)

II. Experimental Considerations for the Beginner

190

(2)

III. Considerations for All

193

(3)

IV. Considerations over the Long Haul

196

(3)

References

199

(5)

11. Cell Line Characterization and Authentication

204

(16)

Joseph Kaplan

Bharati Hukku

I. Introduction

204

(1)

II. Chronology oc Cell Line Authentication Efforrs

205

(1)

III. Markets used for Cell Line Indentification

206

(8)

IV. Conclusion

214

(1)

References

214

(6)

SECTION III Specialilzed Culture Technipues

220

(78)

12. Laboratory Scale-up of Cell Cultures (0.5-50 Liters)

220

(10)

Jennie P. Mather

I. Introduction

220

(1)

II. Roller Bottles

220

(1)

III. Microcarriers

221

(2)

IV. Growing Cells in Hollow Fibers

223

(1)

V. Suspension Adapting Cells

224

(2)

VI. Fermentors

226

(1)

References

227

(3)

13. Cell Synchronization

230

(22)

Gary F. Merrill

I. Introduction

230

(1)

II. Estimating Cell Cycle Phase Durations

231

(6)

III. Synchronizing Cells

237

(9)

IV. Problems in Interpreting Cell Synchrony Experiemnts

246

(2)

References

248

(4)

14. Measurement of Cell Death

252

(14)

Deryk T. Loo

Jill R. Rillema

I. Introduction

252

(1)

II. Cellular Measurement iof Apoptosis

253

(6)

III. Biochemical Measurement of Apoptosis

259

(3)

IV. Concluding Remarks

262

(1)

References

263

(3)

15. Simultaneous Measurement of Cell Cycle and Apoptotic Cell Death

266

(14)

Alison Moore

Christopher J. Donahue

Kenneth D. Donahue

Jennie P. Mather

I. Introdiction

266

(2)

II. Methods

268

(7)

III. Notes

275

(1)

IV. References

276

(4)

16. Embryonic Stem Cells, Creating Transgenic Animals

280

(18)

Maelinda Pirity

Anna-Katerian Hadjantonakis

Andr'as Nagy

I. Introduction

280

(1)

II. Laboratory Equipment

281

(1)

III. Culture Conditions

281

(1)

IV. Maintaining Embryonic Stem (ES) Cells

281

(1)

V. Preparing Tissue Culture Plates for ES Cells

282

(1)

VI. Passaging ES Cells

283

(1)

VII. Freezing ES Cells from Confluent 10-cm Tissue Culture Plates

283

(1)

VIII. Thawing ES Cells from Freezing Vials

284

(1)

IX. Establishing ES Cells

284

(1)

X. Introducing DNA into and Selecting for Genetically Altered ES Cells

285

(1)

XI. Picking ES Cell Colonies

286

(1)

XII. Detecting Genome Alterations

287

(1)

XIII. Freezing ES Cells in 96-Well Plates

287

(1)

XIV. Thawing ES Cells in 96-Well Plates

288

(1)

XV. Chimeras

289

(1)

XVI. Preparation of the Aggregation Plate

289

(1)

XVII. Preparation of Embryos

290

(1)

XVIII. Preparation of Embryos

290

(1)

XIX. ES Cell(-) Embryo Aggregations

291

(1)

XX. Germline Transmission

291

(1)

XXI. Perspectives

292

(1)

References

292

(6)

SECTION IV Microscopy and Morphology

298

(53)

17. Electron Microscopy: Use of Transmission and Scanning Electron Microscopy to Study Cells in Culture

298

(16)

David M. Phillips

I. Introduction

298

(1)

II. Studying Monolayer Cultures awith the Transmission Electron Microscope

298

(6)

III. Cells Grown on Filters or Matrices

304

(1)

IV. Cells Grown in Suspension

305

(3)

V. Studying Monolayer Cultures awith Scanning Electron Morcoscopy

308

(2)

VI. Cells Grown in Suspension

310

(1)

VII. A Few Hints

310

(1)

References

311

(3)

18. Indirect Immunofluorescence Microscopy in Cultured Cells

314

(20)

Sally P. Wheatly

Yu-li Wang

I. Introdiction

314

(1)

II. Cell Culture

314

(1)

III. Sample Preparation for Immunocytochemistry

315

(4)

IV. Antibodies

319

(2)

V. Signal Detection

321

(3)

VI. Miscellaneous Issues

324

(1)

VII. Laboratory Protocols for Fixation Extraction

325

(6)

References

331

(3)

19. Cellular Localization of mRNA and Protein: In Situ Hybridization Histochemistry and in Situ Lilgand Binding

334

(17)

Teresa K. Woodruff

I. Introduction

334

(1)

II. Method: In Situ Hybridization

335

(5)

III. Protocol: In Situ Hybridization

340

(2)

IV. Method: In Situ Ligand Binding

342

(3)

V. Protocol: In Situ Ligand Binding

345

(2)

VI. Analysis of Slides

347

(1)

VII. Combination of in Situ Hybridization with in Situ Ligand Binding, Immunohistochemistry, or Metabolic Studies.

348

(2)

References

350

(1)

Index

351

(12)

Volumes in Series

363

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Methods in Cell Biology: Animal Cell Culture Methods

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Table of Contents

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