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Principles and Practice of Animal Tissue Culture 1st Published,8173715785,9788173715785

Principles and Practice of Animal Tissue Culture 1st Published

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Publisher:Orient Longman Limited, Universities Press (India) Pvt. Ltd.
Published In:2007
ISBN-10:8173715785
ISBN-13:9788173715785
Binding Type:Paperback
Weight:1.06 lbs
Pages:[xiv] + 230 Pages, Illustrations (Partly Col.), Tables, Index, Appendices, References, Abbreviations, Acknowledgements

The Title "Principles and Practice of Animal Tissue Culture 1st Published" is written by Sudha Gangal. This book was published in the year 2007. The ISBN number 8173715785|9788173715785 is assigned to the Paperback version of this title. The book displayed here is a 1st Published edition. This book has total of pp. [xiv] + 230 (Pages). The publisher of this title is Orient Longman Limited, Universities Press (India) Pvt. Ltd.. Principles and Practice of Animal Tissue Culture 1st Published is currently Not Available with us.You can enquire about this book and we will let you know the availability.

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About the Book

The technology of cell, tissue and organ culture is of primary importance in the development of the biotechnology industry. Major advances in the biological sciences have been possible, at least partially, due to this technology. Principles and Practice of Animal Tissue Culture provides basic information the technology of animal tissue culture and discusses several specialized techniques. The Book is well supported by relevant graphs and photographs of equipment and cultured tissues.

The book is a comprehensive practical guide that emphasizes ‘good laboratory practices’. Protocols, aseptic techniques, media preparation and data have been systematically presented to be of help primarily to students of biotechnology.

About the Author

Sudha Gangal is a biomedical scientist with over four decades of experience in research. She is a founder member of the Moving Academy of Medicine and Biomedicine, Pune, and has been the Vice-President of the organization since 2001. Her chief research areas are tumour immunology, cell biology and genetic diseases. She conducts basic and advanced training programmes for students and teachers on a regular basis, and has published several research papers and contributed chapters in several books.

Contents

I. INTRODUCTION :
1. Introduction to Tissue Culture
2. Applications of Tissue Culture
3. Limitations of Tissue Culture

II. DESIGNING THE TISSUE CULTURE LABORATORY :
1. Washing and Sterilization Area
2. Storage Area
3. Sterile Tissue Culture Room

III. EQUIPPING THE TISSUE CULTURE LABORATORY :
1. Equipment Needed for Tissue Culture Laboratory
2. Incubator
3. Autoclave
4. Hot air Oven for Dry Sterilization
5. Refrigerators and-20°C Deep Freezer
6. Inverted Microscope
7. Glassware Washing Facility
8. Water Purification
9. Centrifuge
10. Laminar Flow Benches (Hoods)
11. CO2 Incubator
12. Balances, pH Meter, Vortex Mixer, Magnetic Stirrer
13. Research Microscope
14. Pipetting Devices
15. Media Filtration Assembly
16. Controlled Rate Freezing of Cells and Liquid N2 Storage Facility
17. Hemocytometer

IV. GLASSWARE AND PLASTICWARE USED FOR TISSUE CULTURE :
1. Glassware and Plasticware
2. Substrate Material for Growing Cells
3. Tissue Culture Vessels

V. ASEPTIC TECHNIQUES :
1. Precautions for Maintaining Sterility

VI. STERILIZATION OF MATERIALS TO BE USED FOR TISSUE CULTURE :
1. Items to be Sterilized in the Oven
2. Items to be Sterilized by Autoclaving
3. Sterilization of Filtration Assembly
4. Sterilization of Liquid Reagents and Media
5. Testing Media for Sterility, Toxicity and Growth-promoting Activity
6. Sterilization of Serum

VII. TISSSUE CULTURE MEDIA :
1. Properties and Special Requirements of Media
2. Balanced Salt Solutions
3. Complete Media : Composition of Frequently Used Media
4. Commonly Used Antibiotics
5. Requirement of Serum and Growth Factors
6. Conditioned Medium
7. A note on Serum Free Media

VIII. TYPES OF TISSUE CULTURE :
1. Organ Culture
2. Primary Explant Culture
3. Cell Cultures

IX. ORGAN AND ORGANOTYPIC CULTURES :
1. Organ Cultures
2. Filter Well Inserts
3. Hollow Fibre System
4. Spheroids

X. PRIMARY CULTURES :
1. Practical Notes on Setting up Primary Cultures
2. Epidermal Keratinocytes
3. Corneal Epithelium
4. Mammary Epithelium
5. Cervical Keratinocytes
6. Hepatocytes
7. Pancreatic Epithelium
8. Prostatic Epithelial Cells
9. Chondrocyte Cultures
10. Culture of Osteoblasts
11. Cerebellar Neuronal Cells
12. Glial Cells
13. Melanocyte Cultures
14. Bone Marrow Cultures
15. Culture of Tumour Cells : General Considerations

XI. TRANSFORMATION, DIFFERENTIATION AND DEDIFFERENTIATION :
1. Transformation
2. Differentiation
3. Dedifferentiation

XII. CULTURE OF CELL LINES :
1. Initiation and Evolution of Cell Lines
2. Maintenance of Cell Lines
3. Phases of Cell Cycle
4. Growth Curve of Cells
5. Protocols for Suhculturing of Cells : Monolayers and Suspension Cultures
6. Use of Antibiotics
7. Types of Microbial Contaminants
8. Mycoplasma
9. Viral Contamination
10. Cross Contamination of Cell Lines
11. Setting up Replicate Cultures
12. Cryopreservation of Cells
13. Sources for Seed Cultures

XIII. PROTOCOLS FOR ROUTINE CHARACTERISATION OF CELL LINES :
1. Viable Cell Counting with Hemocytometer
2. Growth Curve : Monolayer Culture
3. Cell Morphology : Giemsa Staining
4. Mitotic Index
5. Cell Cycle Analysis
6. Assessment of Plating Efficiency
7. DNA Estimation and Synthesis
8. Protein Estimation and Synthesis

XIV. SPECIALIZED TECHNIQUES IN TISSUE CULTURE :
1. Cloning : General Information
2. Cloning of Cells in Suspension, Cloning in Agar, Cloning in Methocel
3. Cell separation : Cell Density, Immune Panning, Magnetic Activated Cell Sorting
4. Hematopoietic Cell Colony Forming Assays :
i. Assays for BFU-E, CPU-mix and CFU-GEMM
ii. Assay for CFU-GM

5. Immunofluorescence and Immunoperoxidase Staining
6. Testing of Cytotoxic Drugs :
i. SRB Assay
ii. MTT Assay
iii. XTT Assays

7. Lymphocyte Transformation Assay
8. Transformation of Human B Lymphocytes with EBV
9. Cell Synchronization :
i. Metaphase Block
ii. G2 phase Block
iii. G1/S Phase Block

10. Apoptosis :
i. Staining of Apopotic Cells with PI :
ii. Analysis of DNA Fragmentation

XV. STEM CELLS CULTURES :
1. Stem Cell Biology
2. Therapeutic Cloning
3. Human Embryonal Stem (ES) Cell Culture
4. Human Embryonal Germ Cell (GC) Culture
5. Human Embryonal Carcinoma (Teratocarcinoma) Stem Cell (EC) Cell Culture
6. Potential Uses of Stem Cell Cultures

XVI. AUTORADIOGRAPHY OF CULTURED CELLS :
1. Preparation of Cultures for Autoradiography
2. Technique Using Emulsion
3. Technique Using Kodak AR-IO Stripping Film

XVII. CYTOGENETIC ANALYSIS OF CULTURED CELLS :
1. Chromosome Preparations for Characterization of Cell Lines
2. Whole Blood Culture
3. Bone Marrow Culture
4. Amniotic Cell Culture
5. G Banding Technique
6. High Resolution Banding
7. Fluorescence in Situ Hybridization (FISH) Technique

XVIII. FLOW CYTOMETRIC ANALYSIS OF LYMPHOCYTES AND CULTURED CELLS :
1. Principles of Flow Cytometry
2. Single Colour Analysis (Indirect Method)
3. Dual Colour Analysis
4. DNA Analysis

XIX. HYBRIDOMA CULTURES TO GENERATE MONOCLONAL ANTIBODIES :
1. Limitations of Polyclonal Antibodies
2. Hybridoma Technique : Mouse MAbs
3. Applications of Mouse MAbs and Genetically Engineered MAbs
4. Hybridoma Protocol
5. Maintenance of Mouse Mylemoa Cell Lines
6. Immunization of Mice
7. Fusion Protocol

XX. LASER CONFOCAL MICROSCOPY AND ITS APPLICATION IN CELL Biology :
1. Background
2. History of LCM
3. Basic Principle and Equipment
4. Multiphoton LCM
5. Applications of LCM in Cell Biology : Cellular Organisation and Cellular Dynamics
6. Protocols

XXI. TROUBLESHOOTING :
1. Obvious or Cryptic Contamination
2. Bringing Tissue Specimens into the Tissue Culture Laboratory
3. A Check on Media
4. A Check on Cleaning and Sterilization
5. Maintenance of Cells
6. Cross Contamination of Cell Lines

Answers to Exercises
References

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