In Vitro Bioassay Techniques for Anticancer Drug Discovery and Development

Dhanya Sunil, Ph.D. is an Associate Professor at the Department of Chemistry, Manipal Institute of Technology (MIT), Karnataka, India. She has nine years of research and teaching experience in the field of organic synthesis, medicinal chemistry and anticancer drug development. She has more than thirty journal publications to her credit and has won several best paper awards in conferences both within India and abroad. She is also a recipient of Best Teacher Award from MIT, India. She is a reviewer for reputed journals, such as MedChemComm, Current Drug Metabolism. Pooja. R. Kamath is persuing her final year Ph.D. in Medicinal Chemistry under the guidance of Dr. Dhanya Sunil at the Manipal Institute of Technology (MIT), India. She has completed her post-graduation in chemistry from the National Institute of Technology, Karnataka and worked on a project at Syngene International Limited. She has six research publications to her credit. Dr. Raghu Chandrashekhar is an Associate Professor at the Department of Pharmaceutical Biotechnology, Manipal College of Pharmaceutical Sciences, Manipal. His qualifications include Doctor of Philosophy (PhD). 2007. Dr.MGR Medical University, Chennai, Tamilnadu. M Pharm. 2000-02. Dr.MGR Medical University, Chennai, Tamilnadu - First Class B Pharm. 1992-96. Kuvempu University, Karnataka – Tenth Rank.

Introduction to Anticancer Drug Development.

Cancer.

Chemotherapy.

Need for Effective Anticancer Drugs.

Stages of Anticancer Drug Development.

References.

In Vitro Biological Assays for Anticancer Chemotherapeutics.

Introduction.

Advantages of In Vitro Assays.

Disadvantages of In Vitro Assays.

Classification of In Vitro Assays.

Essential Techniques of Cell Cultures.

Primary Cell Cultures.

Maintenance of Cell Lines.

Subculture of Cells.

References.

Cell Viability and Cytotoxicity Assays.

Introduction.

Cell Membrane Integrity Assays.

Dye Exclusion Assays.

LDH Release Assays.

Functional or Metabolic Assays.

Tetrazolium Reduction Assays.

Sulforhodamine B Assay.

Acid Phosphatase Assay.

Alamar Blue Oxidation-Reduction Assay.

Neutral Red Assay.

Protease Viability Marker Assay.

Cell Adhesion Assay.

References.

Cell Proliferation Assays.

Introduction.

DNA Labeling Assays.

[3H]-Thymidine Assay.

BrdU Incorporation Assay.

Metabolic Assays.

Dye Assays.

ATP Assays.

Clonogenic Reproductive Assay.

References.

Apoptosis Assays.

Introduction.

Light Microscopy.

Western Blot Caspase Activation Assay.

Annexin V Ataining.

DNA Ladder Assay.

ELISA.

TUNEL Assay.

Comet Assay.

Acridine Orange/Ethidium Bromide Staining.

Mitochondrial Membrane Potential Assay.

Chromatin Condensation Assay.

APOPercentageTM Apoptotic Assay.

PARP Cleavage Assay.

Flow Cytometry.

References.

Cell Migratory Assays.

Introduction.

2D Cell Migration Assays.

Wound Healing Assay.

Cell Exclusion Zone Assay.

Transwell Migration Assay.

Fence Assay.

Microfluidic Assays.

Single Cell Motility Assay.

2D/3D Migration Assays.

Microcarrier Bead Assay.

Spheroid Migration Assay.

References.

In Vitro Cell Invasion Assays.

Introduction.

Transwell Invasion Assay.

Platypus Invasion Assay.

Gelatin Degradation Assay.

3D Cell Tracking Assay.

Vertical Gel 3D Invasion Assay.

Spheroid/Monodispersed Cell Invasion Assay.

Spheroid Confrontation Assay.

Spheroid Gel Invasion Assays.

References.

Anti-Angiogenesis Assays.

Introduction.

Endothelial Tube Formation Assay.

Rat Aortic Ring Assay.

References.

High-Throughput Screening Assays.

Introduction.

Cell Cultures.

High-Throughput Cytotoxicity and Apoptosis Screening in 2D Cultures.

3D Cell-Based HTS Assays.

References.

Index.