In Vitro Pharmacology

Oncology


  • India's largest tumor bank repository with human cancer cell lines representing 15 - 20 different cancer types
  • Characterized cell lines from authenticated sources
  • Inclusive of untransformed cell lines to enable early safety assessment
  • Drug resistant cell lines & primary cultures from human biopsies
  • Complete repertoire of viability & cytotoxicity assessment screens
  • Customized Molecular target based screens
  • Combination index studies for assessment of synergy
  • In Vitro Services

    • Cytotoxicity Screens
    • Combination Index
      Model Evaluation of drug interactions in combination cancer chemotherapy by combination index method
      Test system
      • Cancer cell lines
      Methods
        Determination of % cytotoxicity & IC50 of individual drugs Selection of concentrations around IC50 for combination studies Testing all possible combinations of drugs for combined cytotoxicities Calculation of Combination Index (CI)
      • CI=1 - Synergy
      • CI=1 - Additive
      • CI=1 - Antagonism
      End points
      Categorization of drug combinations as synergistic/additive/antagonistic for each combination tested.
      Model DETERMINATION OF SYNERGISTICALLY EFFICACIOUS COMBINATION(S) OF CISPLATIN AND DOXORUBICIN IN HepG-2 CELL LINE IN VITRO BY COMBINATION INDEX METHOD
      Test system
        Hepatocellular carcinoma (HepG-2)
      Methods
      • Cisplatin
      • Doxorubicin
      End points
      Cells were treated at fixed IC50 of one drug (Cisplatin/Doxorubicin) in combination with variable concentrations of the other (Cisplatin/Doxorubicin). MTT assay CI value was calculated to assess the synergistic effect of test items Synergism, additive or antagonism was based on the following criteria:
      • CI = 1 Synergism
      • CI=1 Additive
      • CI=1 Antagonism
      X
    • Drug Uptake
      Model in vivo: model to quantitate intracellular uptake and localization of drugs (e.g. Nanoparticle/liposomal formulations)
      Test system
      • Human tumor cell lines
      • Human normal cells/cell lines
      Methods
      • Treatment of target cell lines with drug
      • Quantitative analysis - Visualization of Cellular uptake, delivery and subcellular distribution of drugs in target cell lines by TEM Transmission Electron Microscopy)/ Fluorescence microscopy
      • Quantitative analysis - Measurement of intracellular concentrations of drug by bioanalytical methods (HPLC and LC/MS)
      End points
      • Intracellular accumulation of drugs
      • Sub cellular distribution/localization
      • Uptake in non target tissues
      • Dose/Time kinetic profiles for drug uptake
      • Correlation with biological activity
      • Uptake/Transport by endocytotic/Non endocytotic routes
      X
    • Angiogenesis Screens
      Model Assessment of anti-cancer activity of the compounds using tube formation assay
      Test system
      • HUVEC Cells
      Methods
      • Seeding of HUVEC cells on matrigel
      • Application of vehicle and compound of interest at appropriate concentrations
      • Validation with Combretastatin A4 or client defined reference standards
      End points
        Quantitative analysis of -
      • Branch area
      • Branch points
      • Tube length
      • Photomicrographs
      X
    • Cell Migration assay
      Model Assessment of anti-cancer activity of the compounds using cell migration assay
      Test system HUVEC Cells
      Method
      • Seeding of HUVEC cells
      • Creation of wound on surface of culture
      • Application of vehicle and Test item at appropriate concentrations
      End points
      • Number of cells migrated to wound area
      X
    • In vitro Angiogenesis model
      Model Assessment of anti-cancer activity of the compounds using tube formation assay
      Test system HUVEC Cells
      Method
      • Seeding of HUVEC cells on matrigel
      • Application of vehicle and compound of interest at appropriate concentrations
      • Validation with Combretastatin A4 or client defined reference standards
      End points
        Quantitative analysis of -
      • Branch area
      • Branch points
      • Tube length
      • Photomicrographs
      X
    • In vitro angiogenesis VEGF
      Model Assessment of VEGF, bFGF & Endostatin
      Test system HUVEC, MDA-MB-231, MCF-7, HCT-116 etc
      Method
      • Seeding of predetermined number of Cell Type
      • Application of vehicle and Test item at appropriate concentrations
      • Incubation for 48 to 72hr
      • Estimation of VEGF, bFGF & endostatin concentration in culture supernatants
      End points
      • % Inhibition
      • IC50 value
      X
    • Apoptosis Screens
      Model Complete repertoire of Screens for assessment of Programmed cell death to enable selection of Proapoptotic & antiapoptotic molecules
      Test system
      • Human normal cells/cell lines
      • Human cancer cell lines
      Apoptotic Pathways
      • Death receptor and mitochondrial pathway
      End points
      • Externalization of Phosphatidyl Serine (PS) on plasma membrane by Annexin-V staining
      • Loss of transmembrane mitochondrial potential by JC-1 staining
      • Quantitation of Reactive oxygen species (ROS)
      • Levels of pro-apoptotic and anti-apoptotic proteins
      • Ras-ERK kinases
      • Caspase-3 activation
      • PARP cleavage
      • DNA fragmentation
      • Micronuclei staining
      • Cell Cycle analysis
      X
    • Mechanistic Profiling
      Model Assessment of anti-leukemic activity of the compounds in SCID mice
      Test system SCID mice
      Models
      • Intravenous inoculation of leukemic cell line
      • Administration of test item and reference item at predetermined time
      • Observation of animals for defined time period
      • Observation of animals in satellite group for occurrence of chloroma
      End points
      • Percentage Change in body weight
      • Survival rate of animals
      • Clinical signs & symptoms
      • Quantification by flow cytometry analysis in bone marrow & peripheral blood
        — Standard surface specific antigen staining - CD45
        — Lineage specific antigen staining
      • Histopathology
      X
    • Target Based Screens
      Core team of Biochemists, Cell biologists & Molecular Biologists to Develop customized client defined target based screens Repertoire of screens
      • Enzyme assays
      • Cell based methods
      • Cell free screens
      • Functional assays
      • Semi-automatic Homogeneous screens for focused targets
        Target Classes
      • Cytoskeletal proteins
      • Enzymes
      • Nuclear receptors
      • Kinases
      • Phosphatases
      • Hormones
      X