Podcast
Questions and Answers
What is a significant limitation of 2D cell culture?
What is a significant limitation of 2D cell culture?
3D cultures have enhanced reliability and predictability of clinical efficacy.
3D cultures have enhanced reliability and predictability of clinical efficacy.
True
What is a multicellular tumor spheroid?
What is a multicellular tumor spheroid?
Sphere-shaped cell colonies that permit growth and functional studies mimicking human tumors.
The __________ method involves seeding cells on non-adhesive surfaces to generate tumor spheroids.
The __________ method involves seeding cells on non-adhesive surfaces to generate tumor spheroids.
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Match the following characteristics with their respective cell culture types:
Match the following characteristics with their respective cell culture types:
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What is a major advantage of 3D cell culture compared to 2D cell culture?
What is a major advantage of 3D cell culture compared to 2D cell culture?
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One limitation of 3D culture is the lack of vascularization.
One limitation of 3D culture is the lack of vascularization.
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Name two methods for generating multicellular tumor spheroids.
Name two methods for generating multicellular tumor spheroids.
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Which of the following describes how tumor spheroids are created using agitation-based approaches?
Which of the following describes how tumor spheroids are created using agitation-based approaches?
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Microfluidic technology allows for precise control over microenvironments affecting cells.
Microfluidic technology allows for precise control over microenvironments affecting cells.
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What are the two types of bioreactors mentioned for growing tumor spheroids through agitation?
What are the two types of bioreactors mentioned for growing tumor spheroids through agitation?
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Tumor spheroids can be generated using highly porous __________ created from natural or synthetic polymers.
Tumor spheroids can be generated using highly porous __________ created from natural or synthetic polymers.
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Match the challenges with their respective descriptions:
Match the challenges with their respective descriptions:
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What is a key challenge when creating multicellular tumor spheroids?
What is a key challenge when creating multicellular tumor spheroids?
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The size and circularity of MCTs are not important metrics for growth assessment.
The size and circularity of MCTs are not important metrics for growth assessment.
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What factors contribute to the variability of spheroids?
What factors contribute to the variability of spheroids?
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What is one limitation of the traditional spheroid forming system?
What is one limitation of the traditional spheroid forming system?
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Microcarriers can be used to decrease the surface area of traditional monolayer cultures.
Microcarriers can be used to decrease the surface area of traditional monolayer cultures.
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Name one application of hollow fiber bioreactors.
Name one application of hollow fiber bioreactors.
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The __________ is a commercialization product that allows for customizable cell interaction studies.
The __________ is a commercialization product that allows for customizable cell interaction studies.
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Match the following types of 3D cultures with their characteristics:
Match the following types of 3D cultures with their characteristics:
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What is a common application of 3D cultures in biomedical research?
What is a common application of 3D cultures in biomedical research?
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Spheroids generated in microfluidic devices are produced more quickly than traditional methods.
Spheroids generated in microfluidic devices are produced more quickly than traditional methods.
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What is a disadvantage of using filter well inserts?
What is a disadvantage of using filter well inserts?
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Study Notes
2D (Monolayer & Suspension) Cell Culture
- Merits: Absolute control of cell environment, easier cell observation, measurement, and manipulation.
- Limitations: Loss of histological organization, polarity, and differentiation; altered gene expression; lack of host tissue microenvironment; increased drug sensitivity.
3D (Organ & Spheroid) Cell Culture
- Merits: Well-characterized to simulate pathophysiological cellular microenvironment, enhanced reliability/predictability of clinical efficacy; minimizes animal studies; reconstitutes tissue-like cyto-architecture with cell-cell and cell-matrix interactions.
- Limitations: Lack of vasculature and host-immune interactions, diffusional transport limitations (O2 and nutrients may not reach all cells, accumulation of toxic wastes),
Multicellular Tumor Spheroids
- Sphere-shaped cell colonies that mimic naturally occurring human tumors.
- Mimics growth and function of normal and malignant tissues.
- Has similar extracellular matrix and cell-cell/matrix interactions to in vivo conditions.
- Contains layers: proliferative, quiescent, necrotic.
Generating Multicellular Tumor Spheroids: Methods
- Liquid overlay: Cells seeded on non-adhesive surfaces, super-hydrophobic agar/agarose is used.
- Hanging drop: Cells seeded in a tray, inverted, suspended via surface tension, and allowed to proliferate.
- Agitation-based: Cells suspended in a container, rotated or stirred.
- Microfluidic technology: Miniaturized devices with chambers for fluid flow, precise control of microenvironments, and high-throughput screening.
- Matrices & scaffolds: Using porous scaffolds like sponge-like structures for arrangement of immobilized 3D spheroids.
Challenges in Multicellular Spheroid Application
- Uniformity and reproducibility: Producing homogeneous shape and size spheroids consistently.
- Assessment: Establishing a valid evaluation method for growth and drug efficacy.
- High-throughput: Developing a high-throughput culture and drug screening platform.
Other 3D Culture Methods
- Microcarriers: Hydrogels used to mimic high surface area-to-volume ratio, allowing anchorage-dependent cells to grow in suspension cultures.
- Filter well inserts: Commercially available system, enabling the study of 3D chemotactic responses and cell interactions.
- Hollow fibers bioreactors: Perfusion chambers using capillary fibers, allows gas and nutrient permeation to support cell growth.
High-Throughput Platforms for Creating MCTs
- Problem: Traditional spheroid generation methods are time-consuming and produce variable-sized spheroids.
- Solution: Spheroids generation in microfluidic devices to address time and size-related issues.
Hollow Fiber Systems
- Ideal for studying synthesis and release of biopharmaceuticals and are now being employed as semi-industrial scale.
Tissue Engineering
- Construction of tissue-equivalent cultures with potential for tissue replacement therapy.
- Includes skin equivalent cultures for burn repair.
3D Cultures' Applications
- Basic cellular processes: Microenvironment modeling for various diseases (cancer, inflammatory toxicity) and studying cellular organization (aggregates, cysts, tubules).
- Drug discovery: Studying drug penetration (cytotoxic drugs, antibodies), the effect of dosages, and interactions with natural barriers.
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Description
Test your knowledge on the key differences between 2D and 3D cell cultures. This quiz covers multicellular tumor spheroids, methods of generation, and advantages of advanced cell cultures. Challenge yourself to match characteristics and identify the limitations and benefits of each culture type.
