3D vs 2D Cell Culture Quiz
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Questions and Answers

What is a significant limitation of 2D cell culture?

  • Cells maintain their histological organization
  • Sufficient cell-cell interactions
  • Altered gene expression and growth characteristics (correct)
  • Enhanced drug resistance

3D cultures have enhanced reliability and predictability of clinical efficacy.

True (A)

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.

<p>liquid overlay</p> Signup and view all the answers

Match the following characteristics with their respective cell culture types:

<p>2D cell culture = Easier observation and manipulation 3D cell culture = Mimics in vivo conditions Multicellular tumor spheroid = Sphere-shaped cell colonies Liquid overlay method = Non-adhesive surfaces</p> Signup and view all the answers

What is a major advantage of 3D cell culture compared to 2D cell culture?

<p>Better representation of tissue microarchitecture (A)</p> Signup and view all the answers

One limitation of 3D culture is the lack of vascularization.

<p>True (A)</p> Signup and view all the answers

Name two methods for generating multicellular tumor spheroids.

<p>Hanging drop method and agitation method.</p> Signup and view all the answers

Which of the following describes how tumor spheroids are created using agitation-based approaches?

<p>Cells experience continuous motion facilitating cell-cell interactions. (D)</p> Signup and view all the answers

Microfluidic technology allows for precise control over microenvironments affecting cells.

<p>True (A)</p> Signup and view all the answers

What are the two types of bioreactors mentioned for growing tumor spheroids through agitation?

<p>Spinner flask bioreactors and rotating cell culture bioreactors.</p> Signup and view all the answers

Tumor spheroids can be generated using highly porous __________ created from natural or synthetic polymers.

<p>scaffolds</p> Signup and view all the answers

Match the challenges with their respective descriptions:

<p>Uniformity &amp; Reproducibility = Consistency in shape and size of MCTs Assessment = Evaluation methods for growth and efficacy High-throughput = Platforms for drug screening Factors affecting variability = Cell type and culture techniques</p> Signup and view all the answers

What is a key challenge when creating multicellular tumor spheroids?

<p>Uniformity and reproducibility. (B)</p> Signup and view all the answers

The size and circularity of MCTs are not important metrics for growth assessment.

<p>False (B)</p> Signup and view all the answers

What factors contribute to the variability of spheroids?

<p>Cell type, culture techniques, medium composition, cell density.</p> Signup and view all the answers

What is one limitation of the traditional spheroid forming system?

<p>It takes a long time to culture and produces spheroids of various sizes. (B)</p> Signup and view all the answers

Microcarriers can be used to decrease the surface area of traditional monolayer cultures.

<p>False (B)</p> Signup and view all the answers

Name one application of hollow fiber bioreactors.

<p>Studying the synthesis and release of biopharmaceuticals.</p> Signup and view all the answers

The __________ is a commercialization product that allows for customizable cell interaction studies.

<p>filter well insert</p> Signup and view all the answers

Match the following types of 3D cultures with their characteristics:

<p>Microcarriers = Hydrogel-based beads for suspension culture Filter well inserts = Contains filters for studying cell interactions Hollow fibers bioreactors = Supports cell growth on outer surface of capillary fibers Spheroids = 3D aggregates of cells formed in culture</p> Signup and view all the answers

What is a common application of 3D cultures in biomedical research?

<p>Studying basic cellular processes (C)</p> Signup and view all the answers

Spheroids generated in microfluidic devices are produced more quickly than traditional methods.

<p>True (A)</p> Signup and view all the answers

What is a disadvantage of using filter well inserts?

<p>They cannot image at high resolution in real-time.</p> Signup and view all the answers

Flashcards

2D cell culture limitation

Cells lose their natural organization, polarity, and differentiation, leading to unreliable results.

Multicellular tumor spheroid

A 3D model that mimics the growth of real tumors, revealing their inner structure and behavior.

Proliferative layer

A layer in a tumor spheroid where cells are dividing rapidly and actively growing.

Quiescent layer

A layer in a tumor spheroid where cells are inactive and not dividing.

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Necrotic layer

A layer in a tumor spheroid where cells have died due to lack of oxygen or nutrients.

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Liquid overlay method

A method to create tumor spheroids by using a non-sticky surface, like special agar, to keep cells from attaching and forcing them to grow as a sphere.

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Hanging drop method

A method to grow tumor spheroids in hanging drops, suspending cells in a liquid droplet, which allows them to form a spherical shape.

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Microfluidic technology

A method to create tumor spheroids by using special devices and microfluidic channels to control and manipulate cell growth and organization.

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ATP assay

A method to assess cell viability by measuring the activity of metabolic enzymes, which are reduced in dead cells.

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Microfluidic Spheroid Generation

A technique utilizing microfluidic devices to efficiently generate spheroids of consistent size and uniform properties.

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Microcarrier 3D Culture

A type of 3D cell culture that utilizes microcarriers, small beads with high surface area, to support cell growth in suspension. Cells can adhere to the microcarriers, forming a 3D environment.

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Filter Well Inserts

A method for studying cell behavior in a 3D environment by culturing cells on a porous membrane. The membrane allows for nutrient exchange and cell growth, mimicking the properties of tissues.

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Hollow Fiber Bioreactors

A bioreactor system using hollow fibers, which are small tubes, to provide a controlled environment for growing cells. Cells adhere to the fibers, forming a 3D structure within the bioreactor.

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Tissue Engineering

A process that involves engineering tissues from cells, often by culturing cells in a 3D environment that mimics the natural structure of the tissue.

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Viability Assay in 3D Cultures

A method for studying cell functions and interactions by analyzing the ability of cells to survive and thrive in a 3D environment.

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3D Cell Imaging

A technique to study 3D cell behavior by observing cell structures and functions in a 3D environment.

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Agitation-based spheroid formation

In this method, the cell suspension is kept in motion, either by gentle stirring or rotation, preventing cells from attaching to the container walls and promoting cell-cell interactions.

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Matrices and scaffolds method

A method of producing tumor spheroids by using highly porous (sponge-like) structures that can be produced from natural or synthetic polymers.

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Uniformity and Reproducibility

A major challenge in spheroid research is ensuring consistent production of spheroids with uniform shape and size, which is crucial for reliable experiments.

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Assessment of MCT

A challenge in spheroid research is determining the best way to assess spheroid growth and drug efficacy, which can be difficult because of their 3D nature.

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High-throughput spheroid culture

A challenge in spheroid research is scaling up the production of spheroids and drug screening, which requires high-throughput technologies and automation.

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Factors affecting spheroid variability

Spheroids consist of cells that interact strongly with each other and their surrounding environment, making it challenging to ensure uniformity and reproducibility in experiments.

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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.

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