Introduction to Cell Culture

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Questions and Answers

What occurs after primary culture reaches confluence?

It requires subculturing. (correct)

It becomes a cell strain.

It enters senescence.

It must undergo transformation.

What is the main characteristic of a finite cell line?

It has a limited number of divisions before senescence. (correct)

It is derived from immortalized cell lines.

It shows extensive genetic changes during passaging.

It can divide indefinitely without senescence.

What happens when a cell line undergoes transformation?

It loses its genetic uniformity.

It turns into a primary culture.

It acquires the ability to divide indefinitely. (correct)

It becomes senescent immediately.

Which term describes a subpopulation of a cell line selected by cloning?

Cell strain Signup and view all the answers

What is the initial process in cell culture?

Isolation and growth of cells Signup and view all the answers

What distinguishes continuous cell lines from finite cell lines?

Continuous cell lines divide indefinitely. Signup and view all the answers

What must happen to cells after they are isolated and before they become a cell line?

They need to reach confluence. Signup and view all the answers

What leads to the genotypic and phenotypic uniformity in a cell line?

Selection of high growth capacity cells during passaging. Signup and view all the answers

Which essential nutrient is NOT typically supplied in a cell culture medium?

Proteins Signup and view all the answers

What is the primary reason most cells must be cultured in an anchoragedependent manner?

They depend on physical attachment for growth and division. Signup and view all the answers

Which type of cells are characterized by having elongated shapes and a bipolar or multipolar arrangement?

Fibroblastic cells Signup and view all the answers

What is the role of DMSO or glycerol in cryopreservation?

To prevent ice crystal formation within cells Signup and view all the answers

Cell culture can provide insights into which of the following aspects of cellular behavior?

The effects of toxic compounds Signup and view all the answers

Which cells are typically characterized by being spherical and grown in suspension?

Lymphoblast-like cells Signup and view all the answers

What advantage does cell culture provide for drug screening and development?

Consistency and reproducibility of results. Signup and view all the answers

Which of the following factors is NOT typically regulated in a cell culture environment?

Cell shape Signup and view all the answers

What is a primary requirement for the layout of a cell culture hood?

It should have items arranged in direct line of sight. Signup and view all the answers

Which statement accurately describes the function of an incubator?

It provides a controlled environment for cell growth. Signup and view all the answers

What should be done to items placed in the cell culture hood before use?

They must be disinfected with 70% ethanol. Signup and view all the answers

What is a crucial feature of humid CO2 incubators compared to dry incubators?

They provide superior control of culture conditions. Signup and view all the answers

How should the workspace in a cell culture hood be maintained?

It must remain clean and uncluttered. Signup and view all the answers

Which of the following is NOT a characteristic of the incubator's design?

Must include a built-in water reservoir. Signup and view all the answers

Which location is suitable for placing a pipettor in the layout of a right-handed cell culture hood?

Front right area. Signup and view all the answers

Why is frequent cleaning important for the incubator?

To prevent contamination of cell cultures. Signup and view all the answers

What materials are suitable for handling in a Class II biosafety cabinet?

BSL-1, 2, and 3 materials Signup and view all the answers

What distinguishes a Class III biosafety cabinet from a Class II?

It is gas-tight and offers the highest level of protection Signup and view all the answers

Which of the following is NOT a characteristic of clean benches?

They provide user protection Signup and view all the answers

How do horizontal flow hoods differ from vertical flow hoods?

They provide air flow toward the user Signup and view all the answers

When should clean benches be avoided?

When handling cell culture materials Signup and view all the answers

What type of air flow do cell culture hoods provide?

Constant, unidirectional flow of HEPA-filtered air Signup and view all the answers

What potential hazards are associated with clean benches?

They can cause cross-contamination Signup and view all the answers

What is a key safety feature of a Class III biosafety cabinet?

It provides the highest level of personnel protection Signup and view all the answers

Study Notes

Cell Culture Introduction

Cell culture involves removing cells from an animal or plant and growing them in a controlled artificial environment
Cells can be directly removed from tissue or derived from existing cell lines/strains
Primary cultures are the initial stage where cells are isolated and grow until they fill the available space (reach confluence)
Subculturing (passaging) is required to provide more space for growth by transferring cells to a new vessel with fresh growth medium

Cell Lines and Strains

A primary culture becomes a cell line after the first subculture
Cell lines derived from primary cultures have a limited lifespan (finite cells)
As cell lines are passaged, cells with higher growth capacity become predominant, leading to more uniform cells
A cell strain is a subpopulation of a cell line positively selected for specific traits through cloning, or other methods
A cell strain may have additional genetic changes from its parent line

Finite vs Continuous Cell Lines

Normal cells divide a limited number of times before losing their ability to grow, this process is called senescence
Finite cell lines are these types of normal cells.
Some cell lines acquire the ability to divide indefinitely after transformation, known as continuous or immortal cell lines
Transformation can be induced chemically or virally, or happen spontaneously

Culture Conditions

Culture conditions vary for different cell types
Essential nutrients, growth factors, hormones, and gases (like O2, CO2) must be included in the growth medium
Appropriate pH, osmotic pressure, and temperature are vital
Most cells are anchorage-dependent and need a solid or semi-solid substrate (e.g., monolayer culture)
Some cultures can be grown in suspension in the medium

Cryopreservation

Surplus cells can be preserved by treating with a protective agent (DMSO or glycerol) and storing them at very low temperatures (-130°C)

Cell Morphology

Cell morphology is categorized by shape and appearance
Fibroblastic cells (or fibroblast-like cells) are bipolar or multipolar with elongated shapes
Epithelial-like cells are polygonal with more regular dimensions, and grow in discrete patches
Lymphoblast-like cells are spherical and grow in suspension

Applications of Cell Culture

Cell culture is important in cellular and molecular biology
Providing excellent model systems for studying normal cell physiology, biochemistry, aging, drug effects, toxicology, mutagenesis, and carcinogenesis
Used in drug screening, development, and large-scale manufacturing of biological compounds (e.g., vaccines, therapeutic proteins)
Consistency and reproducibility of results are main advantages

Cell Culture Lab Safety

Cell culture labs have specific hazards associated with handling cells, tissues, and reagents.
Accidental punctures, spills, chemical exposure, and infectious aerosols are primary concerns
Following biosafety protocols, personal protective equipment (PPE), and aseptic techniques is critical to reduce risk
Four biosafety levels (BSL-1 to BSL-4) differ depending on the agent and hazard of the cell culture

Safety Data Sheets (SDS)

Material Safety Data Sheets (MSDS) provide information on the properties of substances and are used for safe handling
Primary barriers (like biosafety cabinets) minimize exposure to hazardous materials
The use of Personal Protective Equipment (PPE) provides a barrier from hazardous agents which might contaminate the operator.

Sterile Technique

Sterile technique minimizes contamination
Key elements include maintaining a sterile work area, good personal hygiene, and sterile reagents and media.
Steps for aseptic technique are detailed.
Incubator conditions are designed for proper cell growth

Equipment

Standard cell culture equipment includes a laminar flow hood, incubators, centrifuges, refrigerators, and freezers.
Additional equipment for more advanced techniques are also available.

Aseptic Work Areas and Cell Culture Hoods

Cell culture labs need designated work areas restricted to cell culture
Cell culture hoods offer a controlled aseptic environment
This helps to avoid contamination
Class I-III biosafety cabinets provide varied levels of protection for different cell lines' demands.

Clean Benches

Clean benches aren't biosafety cabinets, they mostly protect the product rather than the personnel from contamination.
They are used for non-infectious or less-harmful procedures, but not for cell culturing or drug formulating.

Layouts and Equipment

Hood layout is important to maximize efficiency and minimize contamination, and should be arranged to keep reagents and media at arms reach, with clean work spaces
Freezers, refrigerators, and storage solutions are used to preserve materials (for optimal cell growth conditions, time spent handling cells and materials must be minimized, and contamination avoided)

Incubators

Incubators maintain the appropriate environment for cell growth, with temperature control and air circulation
Different types of incubators exist for varying needs

Storage

Storage areas in cell culture are for media and reagents (liquids), chemicals, specialized equipment, glassware, tissues, and cells.
Storage must be appropriate for the specific needs of the materials and their characteristics, particularly those that are sensitive to light
There are different types of storage, including refrigerators, freezers, and cryogenic storage, used depending on the needs of the experiments
Cell counters are useful instruments for accurately calculating and documenting cell quantities for quantitative growth kinetics

Biological Contamination

Biological and chemical contaminants can compromise cell cultures
Strict aseptic techniques are needed to reduce the frequency and seriousness of contaminations

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Description

This quiz covers the fundamental concepts of cell culture, including primary cultures, cell lines, and strains. You'll learn about the processes of subculturing and the characteristics of finite cells. Perfect for students looking to understand the basics of cell biology and laboratory techniques.