Lecture 17 - In Vitro Stduies Flashcards
(23 cards)
What is the role of In Vitro Assays in drug discovery?
Evaluate safety and efficacy of new natural product drugs
Natural products are often discovered via phenotypic drug discovery, observing therapeutic effects.
How do In Vitro Assays differ from target-based drug discovery?
In Vitro Assays focus on therapeutic effects, while target-based drug discovery focuses on specific molecular targets.
What do regulatory bodies require for natural products (NP) in drug development?
Understanding of mechanism of action.
What is advantageous for NP drug development?
If NP is a single, pure compound.
What are the uses of cell-based assays?
- Mechanism of action studies
- Target discovery and validation
- Treatment response, resistance, or sensitivity
What parameters can be measured in cell-based assays?
- Viability
- Proliferation
- Protein expression
- Gene expression
- Omics approaches
- Genotoxicity and teratogenicity
What methods are used to measure viability in cell-based assays?
- MTT assay for metabolic activity
- Trypan blue for membrane integrity
What are primary cells?
Cells directly from tissue; more physiologically relevant.
What are the disadvantages of using primary cells?
- Expensive
- Limited lifespan
- Sometimes difficult to culture
What are immortalized cell lines?
Cells derived from tumors or genetically modified to proliferate indefinitely.
What are the advantages of immortalized cell lines?
- Easy to grow
- Bankable
- Distributable
What are genetically engineered cell lines used for?
Overexpression or knockout models and target verification.
What are the advantages of 2D cultures?
- High throughput
- Cost-effective
- Automation friendly
What are the disadvantages of 2D cultures?
- Lack of cell-cell and cell-matrix interactions
- Unnatural growth conditions
What are the characteristics of 3D cultures?
- Closer to in vivo environment
- Allow differentiation and tissue-like structures
What is an organoid?
A 3D culture from stem cells forming mini-organs.
What is the advantage of organoids?
Physiologically relevant and personalized.
What is an organ-on-a-chip?
Microfluidic devices mimicking organ environments and function.
What is a principle for disease-relevant assays?
Ensure tissue and genomic context reflects patient/disease state.
What should assay endpoints correlate with?
Clinical outcomes.
What are In Silico Assays?
Computer-based simulations predicting drug interactions and ADMET.
What are the advantages of In Silico Assays?
- Very high throughput
- Cost-effective
What are the limitations of In Silico Assays?
- Depends on quality of input data
- Cannot fully replicate biological complexity
