In Vitro Pharmacology

Metabolic Disease


  • Diabetes
    Model In Vitro Model for evaluation of anti-diabetic activity
    Test system
    • Type I: Insulinoma cell line (RIN5F/INS-1)
    • Type II: 3T3L-1 preadipocytes/adipocytes
    End points
    • Quantitative glucose uptake in adipocytes
    • Cell based assays for inhibition of LPS induced IL-6 release in preadipocytes for insulin resistance
    • Adipolysis assay: Glycerol release in adipocytes
    • Adipogenesis assay: TG accumulation by Oil Red O staining in adipocytes
    • Insulin release assay: insulin secretion in insulinoma cell line
    Model In vitro model for free fatty-acid- (FFA-) induced hepatic steatosis & insulin resistance
    Test system
    • FFA challenged HepG2 cell line
    Modals
    • Identification of non-cytotoxic doses of test item by MTT assay
    • Steatotic challenge with FFA and treatment with test item
    End points
    • Lipid accumulation by Oil Red O staining
    • Quantitative ROS generation by fluorescence.
    • Estimation of pro-inflammatory cytokines by ELISA
    • Quantitative assessment of apoptosis
    Model In vitro model for hepatic fibrosis and cirrhosis
    Test system
    • Rat hepatic stellate cell line (HSC-T6)
    Modals
    • Identification of non-cytotoxic doses of test item by MTT assay
    • Treatment with test item for specific time point
    End points
    • Quantitative estimation of profibrotic markers (collagen and hyaluronic acid)
    • Cell proliferation by MTT assay
    • Quantitation of TGF-ß1 levels by ELISA
    • Quantitative assessment of apoptosis
    X
  • Osteoporosis
    Model Evaluation of anti-osteoporotic potential of test molecule in vitro
    Test system
    • Human osteosarcoma cell line (MG63)
    Methods
    • Treatment of cells with test compounds
    • Determination of osteoblast differentiation through enhanced expression of :
      • Collagen synthesis
      • Alkaline Phosphatase
      • Osteocalcin
      • Osteopontin
      • Bone morphogenetic protein 2
    • Determination of potential to reduce the levels of ROS induced by H2O2
    End points Osteogenic / anti-osteoporotic effects of test molecule.
    Model Evaluation of anti-osteoporotic potential of test molecule in vitro
    Test system
    • Human osteosarcoma cell line (MG63)
    Methods
      Treatment of cells with test compounds
    • Determination of osteoblast differentiation through enhanced expression of
    • Collagen synthesis
    • Alkaline Phosphatase
    • Osteocalcin
    • Osteopontin
    • Bone morphogenetic protein 2
    • Determination of potential to reduce the levels of ROS induced by H2O2
    End points Osteogenic / anti-osteoporotic effects of test molecule.
    X
  • Hypertension & Cardiovascular model
    Model Enzyme assays
    Test system
    • Purified Angiotensin-converting enzyme (ACE)
    • Purified soluble epoxide hydrolase (sEH) Purified Angiotensin-converting enzyme (ACE)
    • Purified soluble epoxide hydrolase (sEH)
    Methods
    • ACE enzyme activity assay with TI and positive control by colorimetric metsEH enzyme activity assay with TI and positive control by fluorescence based method
    End points
    • Inhibition of ACE enzyme activity by test item
    • Inhibition of sEH enzyme activity by test item
    X
  • In vitro model of hepatic steatosis & insulin resistance
    Model In vitro model for free fatty-acid- (FFA-) induced hepatic steatosis & insulin resistance
    Test system
    • FFA challenged HepG2 cell line
    Methods
    • Identification of non-cytotoxic doses of test item by MTT assay
    • Steatotic challenge with FFA and treatment with test item
    End points
    • Lipid accumulation by Oil Red O staining
    • Quantitative ROS generation by fluorescence.
    • Estimation of pro-inflammatory cytokines by ELISA
    • Quantitative assessment of apoptosis
    X
  • In vitro model for hepatic fibrosis
    Model In vitro model for hepatic fibrosis and cirrhosis
    Test system
    • Rat hepatic stellate cell line (HSC-T6
    Methods
    • Identification of non-cytotoxic doses of test item by MTT assay
    • Treatment with test item for specific time point
    End points
    • Quantitative estimation of profibrotic markers (collagen and hyaluronic acid)
    • Cell proliferation by MTT assay
    • Quantitation of TGF-ß1 levels by ELISA
    • Quantitative assessment of apoptosis
    X