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Questions and Answers
When should a new cell line be authenticated?
When should a new cell line be authenticated?
What is NOT a main aspect of aseptic technique?
What is NOT a main aspect of aseptic technique?
Which practice is recommended to maintain aseptic technique?
Which practice is recommended to maintain aseptic technique?
What does the passage number indicate in cell culture?
What does the passage number indicate in cell culture?
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What is the purpose of changing the medium in cell cultures?
What is the purpose of changing the medium in cell cultures?
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Which of the following is an essential step in subculturing cells?
Which of the following is an essential step in subculturing cells?
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What is the primary aim of aseptic technique in cell culture?
What is the primary aim of aseptic technique in cell culture?
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How often should routine quality control checks be performed on active cell cultures?
How often should routine quality control checks be performed on active cell cultures?
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What is the general recommendation regarding the passage range for cells in culture?
What is the general recommendation regarding the passage range for cells in culture?
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What is critical to monitor in cell lines with a finite lifespan during culture?
What is critical to monitor in cell lines with a finite lifespan during culture?
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Which method is NOT typically used for dissociating a confluent monolayer of cells?
Which method is NOT typically used for dissociating a confluent monolayer of cells?
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What is the purpose of controlling the cooling rate during cryopreservation?
What is the purpose of controlling the cooling rate during cryopreservation?
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At what temperature are cells typically stored for long-term storage using liquid nitrogen?
At what temperature are cells typically stored for long-term storage using liquid nitrogen?
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Which of the following materials is essential for cryopreservation?
Which of the following materials is essential for cryopreservation?
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Which factor is least likely to affect cell viability during cryopreservation?
Which factor is least likely to affect cell viability during cryopreservation?
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What is a primary reason for cryopreserving cells?
What is a primary reason for cryopreserving cells?
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What is the significance of the first permanent cell line developed from Rainbow trout in 1962?
What is the significance of the first permanent cell line developed from Rainbow trout in 1962?
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Which Biosafety Level (BSL) is most commonly required for cell culture labs working with human cells?
Which Biosafety Level (BSL) is most commonly required for cell culture labs working with human cells?
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What role does Fetal Bovine Serum (FBS) play in cell culture?
What role does Fetal Bovine Serum (FBS) play in cell culture?
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What equipment is essential for maintaining a sterile working area in cell culture?
What equipment is essential for maintaining a sterile working area in cell culture?
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Which of the following is NOT a common source of cell culture contamination?
Which of the following is NOT a common source of cell culture contamination?
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What is meant by 'sterile technique' in cell culture?
What is meant by 'sterile technique' in cell culture?
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Which ingredient in culture medium maintains osmotic pressure and pH stability?
Which ingredient in culture medium maintains osmotic pressure and pH stability?
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What is a major problem caused by contamination in cell cultures?
What is a major problem caused by contamination in cell cultures?
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How do inverted microscopes differ from regular microscopes?
How do inverted microscopes differ from regular microscopes?
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Which of the following describes Mycoplasma in the context of cell culture contamination?
Which of the following describes Mycoplasma in the context of cell culture contamination?
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What is the function of a 5% CO2 environment in incubators for human cells?
What is the function of a 5% CO2 environment in incubators for human cells?
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What is the minimum Biosafety Level recommended for laboratories working with potentially pathogenic cell lines?
What is the minimum Biosafety Level recommended for laboratories working with potentially pathogenic cell lines?
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Why is the pH indicator phenol red used in culture media?
Why is the pH indicator phenol red used in culture media?
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Which of the following statements about cross-contamination of cell lines is true?
Which of the following statements about cross-contamination of cell lines is true?
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Study Notes
Cell Culture Techniques - Basic Principles
- Cell culture is the removal of cells from an organism and their growth in a favorable artificial environment.
- Organisms used in cell culture include molluscs, fish, rabbits, humans, monkeys, insects, and plants, as well as others.
- Cell culture is used in research (basic, toxicity, drug screening, disease, genetic engineering, and gene therapy), regulatory testing purposes, and industry (protein synthesis, vaccines, tissue/organ replacement, and cellular agriculture).
- Examples of applications in toxicology include studies on heavy metals toxicity and nano-toxicity induced by TiO2 and ZnO.
- Fish cell lines are used in in vitro toxicity testing and to study toxicology pathways.
- Cell lines are used for fish health management (disease diagnosis, safety, nutrition), and research on other challenging aspects of aquaculture.
- Unique collagen fibers are used for biomedical applications. Specific compounds from marine sponge-associated fungi are researched.
- A variety of fish cell lines are available and used in research applications.
How Cells are Obtained
- Primary cultures are directly obtained from the original tissue/organ.
- Cells can be obtained from explant cultures, organ cultures, and organotypic cultures.
- Disaggregated tissue can be used in cell culture, resulting in monolayer formation.
Cell Culture Types
- Primary culture - Cells are obtained from tissue/organ; have a limited number of cell divisions.
- Cell lines - Derived from primary cultures; immortalized (unlimited divisions) or finite (limited divisions).
Primary Culture from Fish
- Fish are anesthetized to isolate cells of interest.
- Liver, gills, and kidneys are common sources for primary cells.
- Cells are isolated using enzymatic solutions, then washed.
- Cell viability is determined.
Cell Immortalization
- This allows cells to divide infinitely.
- It is done through spontaneous mutation or virus infection.
- Some cell lines express Telomerase Reverse Transcriptase (TERT) Protein.
Cell Growth
- Primary cultures have a limited number of cell divisions.
- Finite cell lines also have a limited lifespan, but will divide more times than primary cultures.
- Various cell lines are available for different applications, based on their specific properties (speed, efficiency, cost).
Primary Culture vs Cell Lines
- Primary culture - Retains many differentiated characteristics; higher degree of specificity; more labor intensive.
- Cell lines - Propagated indefinitely; more homogenous in cell type; readily available.
Type of growth
- Adherent cultures - Cells grow attached to a surface.
- Suspension cultures - Cells float freely in the culture medium.
Cell Culture Morphology
- Fibroblastic cells have elongated shapes and grow attached to surfaces.
- Epithelial-like cells are polygonal and grow attached to substrates.
- Lymphoblast-like cells are spherical and usually grow in suspension.
Cell Culture Outputs
- Molecular Biology (gene expression, protein expression)
- Flow Cytometry
- Microscopy
- Spectroscopy
- Cytogenetics
Cell Culture Contaminations
- Chemical contamination - Impurities/toxins in media, serum, and water.
- Biological contamination - Bacteria, mycoplasma, virus, fungi, yeast.
- Cross contamination - Contamination between multiple cell lines in same culture medium.
Cell Culture Authentication
- Methods to confirm identity/origin.
- Karyotyping, Isoenzyme analysis, STR and SNP analysis are used.
- Crucial when using cell cultures for research.
Aseptic Technique
- Goal: to prevent microbial contamination.
- Essential for maintaining high-quality cell culture.
- Includes sterile work areas, reagents, media; proper personal hygiene and handling.
Maintenance of Cell Cultures
- Changing media: maintaining growth conditions.
- Subcultivation: allow propagation.
- Density/Viability tests are done to maintain healthy cell growth
- Freezing/thawing allows for storage of cells.
Subculturing
- Transfer of cells from a previous culture to fresh growth medium for propagation.
- Allows the creation of new cultures from existing cell lines.
Cryopreservation
- Freezing cells for long-term storage.
- Slow/fast cooling rates to minimize risk of ice crystal damage.
- Use of freezing media (DMSO).
- Thawing process to revive.
Co-Culture Models
- Two or more different cell types grown together.
- Mimics complex cellular environments in tissues.
- Useful to study interactions & cell-to-cell communication
3D Cell Culture Technology
- Methods for culturing cells in 3D environments (scaffold-free or scaffold-based).
- This mimics in vivo conditions better than 2D models.
Tumor Microenvironment
- Components that form the environment surrounding a tumor (e.g., immune cells, extracellular matrix, blood vessels).
- 3D models are important to study effects of surrounding tissue(s) on tumor cells.
Cell Culture - 2D vs 3D
- 2D: flat surface, limited cell-cell contact, no gradients, simple setup.
- 3D: more complex, replicates in-vivo conditions, cell-cell contact, gradients.
Cell Culture - Organs-on-chip
- Miniature scale device mimicking organs' functions and microenvironment.
- Possible applications in fish physiology, toxicity testing.
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Description
Explore the essential principles and applications of cell culture techniques used in various fields such as research and industry. This quiz covers the basics of cell removal and growth, types of organisms involved, and specific applications in toxicology and aquaculture. Test your knowledge on the importance of cell culture in modern science.
