Dabur Research Foundation Leader in Metabolic Disease Clinical Trials

preclinical CRO in india

Indian Contract Research Organization

Modules

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

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.

drug discovery and development

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

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

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