75 Questions
What type of stem cells are derived from healthy or diseased tissues?
Tissue-resident stem cells
What is the main advantage of using organoids in cell culture?
High clinical relevance
What type of cell culture is derived from tissues or organs of animals?
Primary cell culture
What are organoids used to create?
Diseases
What type of cell culture is attached to a surface?
Adherent cell culture
What is the difference between primary and secondary cell cultures?
Primary is derived from tissues, secondary is derived from cell lines
What type of stem cells are derived from human embryos?
Human Embryonic stem cells (hES)
What is the purpose of using organoids in cell culture?
To model real-life scenarios
What is the primary focus of cell culture?
Removing cells from an animal or plant and growing them in an artificial environment
What is the term used to describe the culture of a non-disaggregated tissue?
Organ culture
What type of cell culture is characterized by cells growing in a single layer?
Monolayer cell culture
What is the term used to describe cells that have the ability to differentiate into different cell types?
Stem cells
What is the term used to describe a three-dimensional structure composed of cells and extracellular matrix?
Organoid
What type of cell culture is characterized by cells growing in a liquid medium?
Suspended cell culture
What is the term used to describe the study of cells in a controlled environment?
Cell culture
What is the primary advantage of using cell culture models over animal models?
All of the above
What is the primary purpose of maintaining aseptic conditions in cell culture?
To prevent contamination and ensure cell viability
What is the main difference between primary and secondary cell cultures?
Primary cell cultures are derived from tissue samples, while secondary cell cultures are derived from existing cell lines
What is the purpose of cryopreservation in cell culture?
To store cells for long-term preservation
What is a common problem that can arise in cell culture experiments?
Loss of cell characteristics
What is the purpose of cell line authentication?
To verify the identity and purity of cell lines
What is a common contaminant in cell culture experiments?
Bacteria
What is the main advantage of using primary cell cultures over cell lines?
Primary cell cultures are more representative of in vivo models
What is the purpose of immunocytochemistry and immunofluorescence in cell line authentication?
To verify the identity of cell lines
What is a key exemption for cell lines under the Human Tissue Act?
They are not relevant material
What regulations deal with the storage and use of human tissue for human application or therapeutic use?
The Quality and Safety Regulations
What type of culture is not discussed in the provided text?
Stem Cell Culture
What is the primary focus of the 'Quality and Safety Regulations'?
Storage and use of human tissue for human application
What type of cell culture is mentioned in the 'Further Reading' section?
Organoid Culture
What is not required for research using cell lines?
All of the above
What is the focus of the 'Advancements in 3D Cell Culture Systems' article?
Personalizing anti-cancer therapies
What is not subject to the 'Quality and Safety Regulations'?
Primary cell cultures
What is the primary advantage of using primary cell cultures in research?
They provide the most accurate representation of physiology and biochemistry
Why are primary cell cultures not always used in research?
They are too sensitive to changes in the microenvironment
What is the main limitation of using primary cell cultures in labs?
They are impossible to scale up for large-scale experiments
What is the primary difference between primary and secondary cell cultures?
Primary cell cultures are derived from donors, while secondary cell cultures are immortalized
What is the purpose of immortalizing cells in secondary cell culture?
To make them faster to grow and more cost-effective
What is the main advantage of using secondary cell cultures in research?
They are faster to grow and more cost-effective
What is the primary characteristic of primary cell cultures?
They are derived from donors and have a finite life span
What is the main difference between primary cell cultures and stem cells?
Primary cell cultures are derived from donors, while stem cells are derived from embryos
What type of cell culture is characterized by cells growing in a single layer?
2D cell culture
What type of cells are derived from human embryos?
Human Embryonic stem cells (hES)
What is the primary purpose of organoids in cell culture?
To create diseases to get a ‘real-life-like’ scenario
What is the main difference between primary and secondary cell cultures?
Primary cell cultures are derived from tissues or organs, while secondary cell cultures are derived from existing cell lines
What type of cell culture is characterized by cells growing in a three-dimensional structure?
3D cell culture
What is the primary advantage of using organoids in cell culture?
They can be used to create diseases to get a ‘real-life-like’ scenario
What is the primary purpose of maintaining aseptic conditions in cell culture?
To prevent contamination
What is a common contaminant in cell culture experiments?
All of the above
What is the purpose of cryopreservation in cell culture?
To preserve cells for future use
What is the main difference between primary and secondary cell cultures?
Primary cells are from tissues, secondary cells are from cell lines
What is the purpose of cell line authentication?
To verify cell identity
What is a requirement for cell culture?
All of the above
What is a limitation of using primary cell cultures in labs?
They have limited lifespan
What is a method used in cell line authentication?
Immunocytochemistry and Immunofluorescence
What is the main difference between fibroblasts and adipocytes in cell culture?
Function and origin
Which of the following cell types is typically found in neural tissue?
Neurons
What is the key characteristic of monolayer cell culture?
Cells grow in a single layer
What is the main difference between adherent and suspension cell cultures?
Adhesion properties
Which of the following is an example of a cell type that can be found in cell culture?
All of the above
What is the purpose of showing images of different cell types in cell culture?
To illustrate the difference in morphology
What type of tissue is derived from an embryonic chicken?
Medullary plate
Which type of cell culture is characterized by cells that are attached to a surface?
Monolayer
What type of cell culture is characterized by cells that grow in a liquid medium?
Suspended
Which type of cell culture is derived from tissues or organs of animals?
Tissue culture
What type of cell culture is characterized by cells that grow in a three-dimensional structure?
3D
What type of tissue is exempt from the Human Tissue Act?
Cell lines
What type of tissue is subject to the Quality and Safety Regulations?
Cell lines
What type of tissue does not require a HTA licence for storage?
Cell lines
What type of tissue is not subject to ethical review under the Human Tissue Act or NHS research governance systems?
Cell lines
What type of tissue is not subject to the Human Tissue Act?
Cell lines
What type of cells are derived from human embryos and have the ability to differentiate into different cell types?
Human Embryonic stem cells
What type of cells are used to create 'real-life-like' scenarios in cell culture?
Organoids
What type of cells are derived from healthy or diseased tissues and can be used to create diseases in cell culture?
Organoids
What type of cells are derived from tissues or organs of animals and are used in primary cell culture?
Primary cells
What type of cells are used in cell culture to study diseases and are derived from pluripotent or tissue-resident stem cells?
Organoids
Study Notes
Introduction to Cell Culture
- Cell culture refers to the removal of cells from an animal or plant and their subsequent growth in a favorable artificial environment.
- The cells may be removed from the tissue directly and disaggregated by enzymatic or mechanical means before cultivation.
Cell Culture vs Tissue Culture
- "Cell culture" refers to the maintenance in vitro of disaggregated cells.
- "Organ culture" refers to a culture of a non-disaggregated tissue.
- "Tissue culture" encompasses both terms.
Types of Cell Culture
- By origin:
- Tissue specificity (e.g., breast cancer, kidneys, neurons)
- By adhesion:
- Monolayer
- Suspended
- By differentiation:
- Differentiated
- Stem cells
Stages of Cell Culture
- Primary cell culture:
- Tissue sampling
- Isolation of cells
- Washing and purification
- Seeding and expansion
- Cryopreservation (cold storage)
- Secondary cell culture:
Requirements for Cell Culture
- Aseptic environment
- Culture media
- Incubators (CO2, temp, and humidity control)
- Storage facility (liquid nitrogen?)
Contaminants in Cell Culture
- Bacteria
- Yeasts
- Fungi (mold)
- Other mammalian cells
Cell Line Authentication
- Immunocytochemistry and immunofluorescence
- RT-PCR
- Confocal microscopy
- Karyotyping
- Next-generation sequencing
- Flow cytometry
- Western blot
- Immunohistochemistry
Types of Cell Culture - Differentiation
- Human embryonic stem cells (hES)
- Induced pluripotent cells (iPSCs)
Types of Cell Culture - Adhesion
- Monolayer
- Suspended
2D, 3D, and Organoid Cell Cultures
- 2D cell cultures: traditional cell culture method
- 3D cell cultures: more complex and physiologically relevant than 2D cultures
- Organoid cell cultures: simple tissue-engineered cell-based in vitro models
Applications of Cell Culture Technology
- Medicine
- Biotech
- Pharma
Primary vs Secondary Cell Culture
- Primary cell cultures:
- Derived from tissues or organs of animals
- Finite life span
- Take longer to grow and are sensitive to changes in the microenvironment
- Vary greatly in their characteristics depending on the health and genetic state of the donor
- Secondary cell culture - cell lines:
- Initially derived from a donor animal/organ
- Immortalized artificially or naturally
- Faster growth
- Well-established conditions and protocols
- Lower cost
Cell Culture
- Cell culture refers to the removal of cells from an animal or plant and their subsequent growth in a favorable artificial environment.
- Cells may be removed from the tissue directly and disaggregated by enzymatic or mechanical means before cultivation, or they may be derived from a cell line or cell strain that has already been established.
Types of Cell Culture
- By Origin:
- Tissue specificity (e.g. breast cancer, kidneys, neurons)
- By Adhesion:
- Monolayer
- Suspended
- By Differentiation:
- Differentiated
- Stem Cells
2D vs 3D Cell Culture
- 2D: monolayer cell culture
- 3D: cell culture in three-dimensional structures, such as spheroids or organoids
Organoids
- Simple tissue-engineered cell-based in vitro models derived from pluripotent or tissue-resident stem cells, or progenitor or differentiated cells from healthy or diseased tissues.
- Used to create diseases to get a ‘real-life-like’ scenario.
Primary vs Secondary Cell Culture
- Primary cell cultures: derived from tissues or organs of animals.
- Secondary cell cultures: derived from cell lines or cell strains that have already been established.
Stages of Cell Culture
- Primary cell culture:
- Tissue sampling
- Isolation of cells
- Washing and purification
- Seeding and expansion
- Cryopreservation (cold storage)
- Secondary cell culture: similar stages, but with the use of existing cell lines.
Requirements for Cell Culture
- Aseptic environment
- Culture media
- Incubators with CO2, temperature, and humidity control
- Storage facility (liquid nitrogen)
Contamination and Cell Line Authentication
- Contamination risks: bacteria, yeasts, fungi, and other mammalian cells.
- Cell line authentication methods:
- Immunocytochemistry and immunofluorescence
- RT-PCR
- Confocal microscopy
- Karyotyping
- Next-generation sequencing
- Flow cytometry
- Western blot
- Immunohistochemistry
Introduction to Cell Culture
- Cell culture refers to the removal of cells from an animal or plant and their subsequent growth in a favorable artificial environment.
- Cells may be removed from tissue directly and disaggregated by enzymatic or mechanical means before cultivation, or they may be derived from a cell line or cell strain that has already been established.
Cell Culture vs Tissue Culture
- "Cell culture" refers to the maintenance in vitro of disaggregated cells.
- "Organ culture" refers to a culture of a non-disaggregated tissue.
- "Tissue culture" encompasses both cell culture and organ culture.
Types of Cell Culture
- By Origin:
- Tissue specificity (Breast cancer, Kidneys, Neurons)
- By Adhesion:
- Monolayer
- Suspended
- By Differentiation:
- Differentiated
- Stem Cells
Cell Lines
- Cell lines are not relevant material under the Human Tissue Act.
- Storage of cell lines for research does not require a HTA licence.
- Cell lines are subject to the Human Tissue (Quality and Safety for Human Application) Regulations 2007.
- Research using cell lines does not require ethical review under the Human Tissue Act or NHS research governance systems.
Application of Cell Culture Technology
- Primary Cell culture applications:
- Cancer research
- Vaccine development
- In vitro toxicology testing
- Pharmaceutical testing
2D and 3D Cell Culture
- 2D cell culture: cells grow in a flat, two-dimensional structure.
- 3D cell culture: cells grow in a three-dimensional structure, mimicking in vivo conditions.
Organoids
- Derived from pluripotent or tissue-resident stem cells from healthy or diseased tissues, such as tumours.
- Used to create diseases to get a ‘real-life-like’ scenario.
- Components of organoid cultures:
- ECM components
- Growth factors
- Small molecules
- Matrices
Primary vs Secondary Cell Culture
- Primary cell cultures: derived from tissues or organs of animals.
- Secondary cell cultures: derived from primary cell cultures or cell lines.
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