| Overview |
|
Fibrosis, characterized by tissue scarring, is usually chronic and progressive as clinical symptom. It could be caused by diseases, iatrogenic injury or trauma. Those fibrosis-associated diseases include cirrhosis (LF), hepatitis, nonalcoholic steatohepatitis (NASH), chronic kidney disease (CKD), myocardial infarction, heart failure, diabetes, celiac disease, scleroderma and idiopathic pulmonary fibrosis (IPF), etc. The relevant abnormalities affect about one fourth of population in world, but with no such effective drugs available until |
now. |
|
| Fibrosis treatment options |
• |
|
Suppression of injury stimuli |
• |
Anti-inflammation |
• |
Inhibition of tissue specific cell activation (i.e. lung fibroblast; hepatic stellate cells) |
• |
Prevention of new ECM accumulation, etc. |
|
|
 |
|
The establishment and application of fibrosis disease models related to human diseases have attracted more and more industry’s attention and become an indispensable part in pathogenesis research and therapeutic drug discovery. Here at HDB, we offer a comprehensive package for preclinical fibrosis drug discovery in both In-vitro assay development & screen to In-vivo disease models. Find out how we can accelerate your |
R&D effort for the treatment of fibrosis diseases.
|
|
| Markers of organ-specific In-vitro fibrosis models
|
|
 |
|
| Selected In Vitro Assay List for Fibrosis Drug Discovery
|
|
| Cell Based qRT-PCR based Fibrosis Model |
• |
|
Compound tests for anti-fibrotic activity in TGF-β induced MRC-5 cells by RT-PCR |
• |
Detection of expression of ECM related genes in Traditional Chinese Medicine (TCM) F6 |
|
treated LX-2 cells by RT-PCR |
• |
Compound test for anti-fibrotic activity in TGF-β induced MRC-5 cells by Collagen |
|
protein level analysis |
|
|
| Cell-based ELISA and WB for Fibro-genesis Related Targets |
• |
|
Detection of expression of α-SMA in TCM treated LX-2 cells by ELISA |
• |
Expression of α-SMA, collagen I, collagen III in TCM treated LX-2 cells (WB) |
|
|
| Cell-Image Based High Content Assay for Fibrosis |
• |
|
Validated assay in Anti-renal Fibrosis in TGF-β1 activated NRK47F cells (HCS) |
|
|
| Apoptosis and Binding Assays |
• |
|
Apoptosis Assay |
• |
Binding assay of AbX to PDGF-BB (Biacore) |
|
|
|
|
| Assay #1: Compound tests for anti-fibrotic activity in TGF-β induced MRC-5 cells by RT-PCR |
|
Quantitative RT-PCR assay development: |
Selected target genes > Sample preparation > cDNA synthesis > Optimization of RT-PCR |
 |
| Purpose: To test the in vitro concentration-response relationship of the anti-fibrotic activity of test compounds by measuring the inhibition of TGF-β induced α-SMA mRNA expression in human lung fibroblast cells
|
|
|
|
| Assay #2: Detection of expression of ECM related genes in Traditional Chinese Medicine (TCM) F6 treated LX-2 cells by RT-PCR |
|
 |
| Compound F6 showed effective on the expression of ECM related genes in F6 treated LX-2 cells
|
|
|
|
| Assay #3: Compound test for anti-fibrotic activity in TGF-β induced MRC-5 cells by Collagen protein level analysis |
|
 |
| The recovered cell culture supernatants after 48 h of TGF-β/test compound treatment were subjected to Sircol assay to analyze collagen protein levels |
|
|
|
| Assay #4: Detection of expression of α-SMA in TCM treated LX-2 cells by ELISA |
|
 |
| Compound 861 and F6 (Fraction from Traditional Chinese Medicine) showed inhibition on the expression of α-SMA in LX-2 cells |
|
|
|
| Assay #5: Expression of α-SMA, collagen I, collagen III in TCM treated LX-2 cells (WB) |
|
 |
| Compound 861 and its components showed inhibition on the expression of α-SMA and collagens in TCM treated LX-2 cells |
|
|
|
| Assay #6: Validated assay in Anti-renal Fibrosis in TGF-β1 activated NRK47F cells (HCS) |
|
 |
| NRK49F cells were seeded to 96-well plate. The cells were activated by TGF-β1, a major pro-fibrosis factor, and induced to myofibroblasts, which was the primary sources of ECM and characterized by over-expression of α-SMA. Through the test and analysis of the expression of α-SMA of cells incubated with different concentration of curcumin, it can be found that curcumin can inhibit the activation of NRK49F cells induced by TGF-β1.
|
|
|
|
| Assay #7: Apoptosis Assay |
|
| Purpose: To develop apoptosis assay to evaluate the compound/antibody effect on FasL-induced apoptosis which causes liver injury
|
 |
• |
|
Cells: Jurkat cells, PBMC |
• |
|
Selected reagent: Caspase 3/7 |
| Assay optimization: Cell density, Incubation time |
|
|
|
|
|
|
| Assay #8: Binding assay of AbX to PDGF-BB (Biacore) |
|
| Instrument: Biacore 3000 |
• |
|
Sensor chip: CM5 |
• |
Assay optimization: Chip Coating |
|
| Kinetic analysis of AbX binding to PDGF-BB by Biacore |
 |
|
|
|
| Summary |
Over the years, HDB has established a variety of in vitro fibrosis assays ranging from gene expression to protein expression assays, further to phenotypic and MOA assays. The cell-based fibrosis models for Pulmonary (Lung) fibrosis, Hepatic (Liver) fibrosis, Renal (Kidney) fibrosis were developed and successfully performed on routine base, helping various |
clients with their research and drug discovery programs. |
|
| Find more about HDB’s comprehensive collection of Fibrosis In vivo Models >>> |
|
沪公网安备 31011502003320号