In Vitro Biological Systems (eBook)

Front Cover 1
Methods in Toxicology 4
Copyright Page 5
Table of Contents 6
Contributors 12
Preface 18
Introduction 20
References 22
SECTION I: NEURAL AND NEUROMUSCULAR SYSTEMS 24
CHAPTER 1. HIPPOCAMPAL SLICES 24
INTRODUCTION 24
MATERIALS 24
PROCEDURES 27
COMMENTS 28
REFERENCES 29
CHAPTER 2. OGANOTYPIC NEURAL CULTURES 30
INTRODUCTION 30
MATERIALS AND REAGENTS 31
PROCEDURES 34
COMMENTS 42
ACKNOWLEDGMENTS 48
REFERENCES 48
CHAPTER 3. REAGGREGATE CULTURES FOR NEUROTOXICOLOGICAL STUDIES 50
INTRODUCTION 50
MATERIALS AND REAGENTS 52
PROCEDURES 54
EVALUATION OF NEUROTOXICITY 57
EXPERIMENTAL APPROACHES 63
COMMENTS 66
REFERENCES 67
CHAPTER 4. CYTOTOXICITY IN MURINE NEOCORTICAL CELL CULTURE 69
INTRODUCTION 69
CULTURE MEDIA AND SOLUTIONS 69
CULTURE PROCEDURES 71
EXPERIMENTAL PROTOCOLS 76
DISCUSSION 81
ACKNOWLEDGMENTS 83
REFERENCES 83
CHAPTER 5. CULTURES OF ADULT TRIGEMINAL GANGLION NEURONS 84
INTRODUCTION 84
MATERIALS AND REAGENTS 84
PROCEDURES 86
COMMENTS 90
ACKNOWLEGMENTS 91
REFERENCES 91
CHAPTER 6. ADRENOMEDULLARY CHROMAFFIN CELLS FOR TOXICOLOGICAL STUDIES 93
INTRODUCTION 93
MATERIALS AND REAGENTS 95
PROCEDURES 97
COMMENTS 99
ACKNOWLEDGMENTS 103
REFERENCES 103
CHAPTER 7. PC-12 CELLS 105
INTRODUCTION 105
MATERIALS AND REAGENTS 105
PROCEDURES 106
COMMENTS 114
ACKNOWLEDGMENTS 116
REFERENCES 116
SECTION II: OCULAR SYSTEM 117
CHAPTER 8. CORNEAL EPITHELIAL AND ENDOTHELIAL CELL CULTURE 117
INTRODUCTION 117
RABBIT CORNEAL EPITHELIAL CELL CULTURE 118
RABBIT CORNEAL ENDOTHELIAL CELL CULTURE 120
MODELS OF ENDOTHELIAL DAMAGE 122
ACKNOWLEDGMENTS 122
REFERENCES 122
CHAPTER 9. ISOLATION AND CULTURE OF RETINAL EPITHELIUM 123
INTRODUCTION 123
HUMAN RETINAL PIGMENT EPITHELIUM 124
SUBCULTURING HUMAN RETINAL PIGMENT EPITHELIUM 128
COMMENTS 129
OTHER MAMMALIAN RETINAL PIGMENT EPITHELIA 132
MYOMORPHIC RETINAL PIGMENT EPITHELIUM 132
REFERENCES 134
SECTION III: RESPIRATORY SYSTEM 135
CHAPTER 10. ISOLATION AND CULTURE OF TYPE II ALVEOLAR EPITHELIAL CELLS 135
INTRODUCTION 135
ISOLATION OF TYPE II CELLS 135
TYPE II CELL CULTURES 140
CONCLUSIONS 141
APPENDIX: ISOLATION OF TYPE II CELLS FROM RABBIT LUNG 142
ACKNOWLEDGMENTS 145
REFERENCES 145
CHAPTER 11. ISOLATION OF NONCILIATED BRONCHIOLAR (CLARA) EPITHELIAL CELL FROM MOUSE LUNG 148
INTRODUCTION 148
MATERIALS AND REAGENTS 150
PROCEDURE 151
COMMENTS 154
ACKNOWLEDGMENTS 156
REFERENCES 157
CHAPTER 12. RAT TRACHEAL EPITHELIAL CELL CULTURES FOR STUDIES IN TOXICOLOGY AND CARCINOGENICITY 159
INTRODUCTION 159
MATERIALS AND METHODS 159
SUMMARY AND CONCLUSION 168
REFERENCES 170
SECTION IV: CARDIOVASCULAR SYSTEM 172
CHAPTER 13. PREPARATION OF PRIMARY CULTURES OF POSTNATAL RAT MYOCARDIAL CELLS FOR TOXICOLOGICAL STUDIES 172
INTRODUCTION 172
MATERIALS AND REAGENTS 173
PROCEDURES 174
COMMENTS 177
ACKNOWLEDGMENTS 183
REFERENCES 183
CHAPTER 14. AORTIC ENDOTHELIAL AND SMOOTH MUSCLE CELL CULTURES 184
INTRODUCTION 184
MATERIALS AND REAGENTS 185
PROCEDURE 186
COMMENTS 190
ACKNOWLEDGMENTS 192
REFERENCES 192
CHAPTER 15. VESSEL CYLINDERS 194
INTRODUCTION 194
PROCEDURES, MATERIALS, AND REAGENTS 196
COMMENTS 205
REFERENCES 205
SECTION V: GASTROINTESTINAL SYSTEM 207
CHAPTER 16. GASTRIC MUCOSAL CELL CULTURE FOR TOXICOLOGICAL STUDY 207
INTRODUCTION 207
MATERIALS AND REAGENTS 208
PROCEDURE 208
MORPHOLOGICAL STUDIES 209
CELLULAR FUNCTIONS 211
COMMENTS 212
ACKNOWLEDGMENTS 215
REFERENCES 216
CHAPTER 17. SMALL INTESTINAL ENTEROCYTES 218
INTRODUCTION 218
MATERIALS 220
PROCEDURES 221
COMMENTS 222
ACKNOWLEDGMENTS 225
REFERENCES 225
CHAPTER 18. MAINTENANCE AND CHARACTERIZATION OF AN ORGAN CULTURE SYSTEM FOR RAT COLONIC MUCOSA 227
INTRODUCTION 227
MATERIALS AND REAGENTS 227
PROCEDURE 228
RESULTS 231
COMMENTS 243
ACKNOWLEDGMENTS 245
REFERENCES 245
SECTION VIA: LIVER SLICES 247
CHAPTER 19. PRECISION-CUT RAT LIVER SLICES IN DYNAMIC ORGAN CULTURE FOR STRUCTURE–TOXICITY STUDIES 247
INTRODUCTION 247
TISSUE SLICE TECHNOLOGY 248
COMMENTS 250
ACKNOWLEDGMENTS 254
REFERENCES 254
SECTION VIB: LIVER: HEPATOCYTES 256
CHAPTER 20. ISOLATION OF HEPATOCYTES BY COLLAGENASE PERFUSION 256
INTRODUCTION 256
MATERIALS AND REAGENTS 257
PROCEDURE 260
COMMENTS 263
REFERENCES 267
CHAPTER 21. PREPARATION OF PRIMARY MONOLAYER CULTURES OF POSTNATAL RAT LIVER CELLS FOR HEPATOTOXIC ASSESSMENT OF XENOBIOTICS 269
INTRODUCTION 269
MATERIALS AND REAGENTS 270
PROCEDURES 273
COMMENTS 275
REFERENCES 279
CHAPTER 22. ISOLATION AND CULTURE OF HEPATOCYTES FROM DIFFERENT LABORATORY SPECIES 280
INTRODUCTION 280
MATERIALS AND REAGENTS 281
PROCEDURE 282
CULTURE 284
COMMENTS 284
REFERENCES 285
CHAPTER 23. ISOLATION OF HUMAN HEPATOCYTES BY BIOPSY PERFUSION METHODS 287
INTRODUCTION 287
MATERIALS AND REAGENTS 288
PROCEDURE 290
COMMENTS 293
ACKNOWLEDGMENTS 294
REFERENCES 294
CHAPTER 24. HUMAN HEPATOCYTE CULTURES 296
INTRODUCTION 296
REAGENTS AND MATERIALS 296
PROCEDURES 297
COMMENTS 299
REFERENCES 302
SECTION VIC: LIVER: OTHER CELL SYSTEMS 304
CHAPTER 25. HEPATOCYTE AND SPLEEN CELL SYSTEMS 304
INTRODUCTION 304
MATERIALS AND METHODS 306
PROCEDURES 308
ADDITIONAL COMMENTS 313
ACKNOWLEDGMENTS 315
REFERENCES 315
CHAPTER 26. ISOLATION AND CULTURE OF HEPATIC NONPARENCHYMAL CELLS 317
INTRODUCTION 317
EQUIPMENT REQUIRED 318
MATERIALS AND REAGENTS 319
PROCEDURES 321
COMMENTS 327
ACKNOWLEDGMENTS 334
REFERENCES 334
CHAPTER 27. PROLIFERATING LINES OF RAT LIVER EPITHELIAL CELLS 336
INTRODUCTION 336
MATERIALS AND REAGENTS 336
METHOD 339
APPLICATIONS TO TOXICOLOGY 340
ACKNOWLEDGMENTS 341
REFERENCES 341
CHAPTER 28. ISOLATION AND CULTURE OF LIVER EPITHELIAL CELLS FROM CARCINOGEN- TREATED RATS 342
INTRODUCTION 342
GENERAL PROCEDURES 342
LIVER PERFUSION 343
SECONDARY ENZYMATIC DIGESTION 346
CENTRIFUGAL ELUTRIATION 349
CELL CULTURE OF ISOLATED LIVER EPITHELIAL CELLS 352
REFERENCES 354
SECTION VIIA: KIDNEY: PROXIMAL TUBULE FRAGMENTS 355
CHAPTER 29. RENAL PROXIMAL TUBULE PREPARATIONS ISOLATED FROM RAT OR RABBIT KIDNEYS WITHOUT THE USE OF COLLAGENASE 355
INTRODUCTION 355
MATERIALS AND METHODS 356
COMMENTS 358
ACKNOWLEDGMENTS 363
REFERENCES 363
CHAPTER 30. SUSPENSION CULTURE OF RABBIT RENAL PROXIMAL TUBULES 364
INTRODUCTION 364
MATERIALS AND REAGENTS 365
PROCEDURES 366
COMMENTS 370
REFERENCES 371
CHAPTER 31. PERFUSION METHOD FOR ISOLATION OF RENAL PROXIMAL TUBULES FROM MAMMALIAN SPECIES 373
INTRODUCTION 373
MATERIALS AND REAGENTS 374
PROCEDURE 376
COMMENTS 379
ACKNOWLEDGMENTS 381
REFERENCES 381
CHAPTER 32. ISOLATION AND MAINTENANCE OF MILLIGRAM QUANTITIES OF RABBIT RENAL PROXIMAL STRAIGHT AND CONVOLUTED TUBULES 382
INTRODUCTION 382
MATERIALS AND REAGENTS 382
PROCEDURE FOR ISOLATING AND MAINTAINING STRAIGHT AND CONVOLUTED PROXIMAL TUBULES 384
COMMENTS 387
REFERENCES 390
SECTION VIIB: KIDNEY CELL CULTURE 391
CHAPTER 33. RABBIT CORTICAL EPITHELIAL CELLS 391
INTRODUCTION 391
MATERIALS AND REAGENTS 392
PROCEDURES 393
COMMENTS 396
REFERENCES 397
CHAPTER 34. CULTURE OF RAT KIDNEY PROXIMAL TUBULE EPITHELIAL CELLS IN DEFINED MEDIUM 399
INTRODUCTION 399
MATERIALS AND REAGENTS 400
PROCEDURES 401
COMMENTS 406
ACKNOWLEDGMENTS 408
REFERENCES 408
CHAPTER 35. PRIMARY CULTURE OF RAT RENAL MEDULLARY INTERSTITIAL CELLS 410
INTRODUCTION 410
MATERIALS AND REAGENTS 410
PROCEDURES 411
COMMENTS 416
REFERENCES 420
CHAPTER 36. PURIFICATION OF RENAL CORTICAL CELL POPULATIONS BY PERCOLL DENSITY-GRADIENT CENTRIFUGATION 422
INTRODUCTION 422
MATERIALS AND REAGENTS 423
EXPERIMENTAL PROCEDURES 425
COMMENTS 432
ACKNOWLEDGMENTS 435
REFERENCES 435
CHAPTER 37. RENAL CELL LINES DERIVED FROM RABBIT AND PIG KIDNEY 436
INTRODUCTION 436
MATERIALS AND REAGENTS 437
PROCEDURES 438
COMMENTS 441
REFERENCES 443
SECTION VIII: REPRODUCTIVE AND DEVELOPMENTAL SYSTEMS 445
CHAPTER 38. WHOLE EMBRYO CULTURE: ORGANOGENESIS OF RODENT EMBRYOS IN VITRO 445
INTRODUCTION 445
MATERIALS 445
PREPARATION AND MAINTENANCE OF EMBRYOS 448
METHODS OF ASSESSMENT 453
REFERENCES 455
CHAPTER 39. PREPARATION AND USE OF SERTOLI–GERM CELL COEULTURES FROM 28-DAY-OLD RATS 456
INTRODUCTION 456
MATERIALS AND REAGENTS 458
PREPARATION PROCEDURE 460
PROTOCOL 460
MAINTAINING CULTURES 470
SELECTION AND DETERMINATION OF TOXICANT-ASSOCIATED END POINTS IN COCULTURES 473
ACKNOWLEDGMENTS 476
REFERENCES 476
SECTION IX: IMMUNE SYSTEM 480
CHAPTER 40. ISOLATION AND IN VITRO CULTURE OF MONONUCLEAR PHAGOCYTES 480
INTRODUCTION 480
ISOLATION OF MONONUCLEAR PHAGOCYTES 482
IN VITRO CULTURE AND MACROPHAGE ACTIVATION 487
IDENTIFICATION OF MACROPHAGES WITH NONSPECIFIC ESTERASE STAIN 489
REFERENCES 491
CHAPTER 41. T-LYMPHOCYTE ASSAYS 492
INTRODUCTION 492
MATERIALS AND PROCEDURES 494
REFERENCES 506
CHAPTER 42. LYMPHOCYTES: COMPARISON OF MURINE AND HUMAN B-CELL IMMUNOCOMPETENCE 507
INTRODUCTION 507
MATERIALS AND METHODS 508
ADDITIONAL COMMENTS 515
ACKNOWLEDGMENTS 515
REFERENCES 515
CHAPTER 43. BONE MARROW STROMAL CELL CULTURES 517
INTRODUCTION 517
MATERIALS AND METHODS 518
SUMMARY 526
REFERENCES 528
SECTION X: SKIN 529
CHAPTER 44. PENETRATION STUDIES WITH EXCISED HUMAN AND ANIMAL SKIN 529
INTRODUCTION 529
MATERIALS AND REAGENTS 530
PROCEDURES 535
COMMENTS 538
ACKNOWLEDGMENTS 538
REFERENCES 538
CHAPTER 45. PORCINE SKIN FLAPS 540
INTRODUCTION 540
MATERIALS AND PROCEDURES 541
COMMENTS 547
REFERENCES 548
CHAPTER 46. KERATINOCYTES 550
INTRODUCTION 550
MATERIALS AND REAGENTS 551
PROCEDURES 552
CONCLUSION 554
REFERENCES 554
Index 556