Cell Culture Applications and Techniques Quiz

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Questions and Answers

What are the three applications of cell culture?

The three applications of cell culture are: 1) studying cell physiology and biochemistry, 2) producing biopharmaceuticals, and 3) regenerative medicine and tissue engineering.

How do primary culture and secondary culture differ?

Primary culture is derived directly from a tissue or organ, while secondary culture is derived from an established primary culture.

What are the steps involved in using cryopreservation for cells?

The steps involved in using cryopreservation for cells include: 1) preparing the cryoprotectant solution, 2) suspending the cells in the cryoprotectant, 3) freezing the cells gradually, and 4) storing the frozen cells in liquid nitrogen.

Why do we use subculture for cells, and what are the steps for subculture (passaging)?

Subculture is used to maintain and expand the cell population. The steps for subculture involve: 1) detaching the cells from the culture vessel, 2) counting the cells, 3) seeding the appropriate number of cells in a new vessel, and 4) providing fresh medium for growth. Signup and view all the answers

What are the challenges in DNA purification, and how can endogenous nucleases be effectively inactivated?

The challenges in DNA purification include separating DNA from other cellular components and avoiding fragmentation of long DNA molecules. Endogenous nucleases can be effectively inactivated by using DNase enzymes and preventing them from digesting the genomic DNA. Signup and view all the answers

Define cell culture, cell strainer, anchorage-dependent, and anchorage-independent.

Cell culture is the process of growing cells outside of their natural environment. A cell strainer is a device used to separate cells from a suspension. Anchorage-dependent cells require a surface for attachment and growth, while anchorage-independent cells can grow in suspension. Signup and view all the answers

What are the three basic steps in DNA extraction?

<ol> <li>Cells collection and breaking the cell membranes to expose the DNA, 2. Breaking proteins and RNA, 3. DNA purification</li> </ol> Signup and view all the answers

What is the purpose of adding a protease in DNA extraction?

To break down proteins (optional) Signup and view all the answers

How is debris such as broken proteins, lipids, and RNA treated in DNA extraction?

Treated with concentrated salt solution to make them clump together Signup and view all the answers

What is the purpose of centrifugation in DNA extraction?

To separate the clumped cellular debris from the DNA Signup and view all the answers

What is the purpose of ethanol precipitation in DNA extraction?

To purify the DNA from detergents, proteins, salts, and reagents used during cell lysis Signup and view all the answers

How is DNA isolated in the minicolumn purification step?

Nucleic acids bind to the solid phase (silica or other) depending on the pH and the salt concentration of the buffer Signup and view all the answers

What is the traditional method used for genomic DNA isolation?

Phenol extraction Signup and view all the answers

What is the purpose of isoamyl alcohol in genomic DNA isolation using phenol extraction?

To prevent foaming Signup and view all the answers

How are proteins removed in the separation by salting out method?

Proteins are dehydrated, lose solubility, and precipitate at high salt concentration, then removed by centrifugation Signup and view all the answers

What happens to DNA during separation by salting out?

DNA remains in the supernatant Signup and view all the answers

Who suggested the name 'stem cells' for the cells which can differentiate into another cell type?

Alexander Maksimov Signup and view all the answers

In what year were embryonic stem cells (ES cells) first derived from mouse embryos?

1981 Signup and view all the answers

Who discovered mesenchymal stem cells in 1991?

Arnold Kaplan Signup and view all the answers

Who were the two groups that independently first derived embryonic stem cells (ES cells) from mouse embryos?

1-Martin Evans and Matthew Kaufman from the Department of Genetics, University of Cambridge, 2-Gail R. Martin, University of California, San Francisco Signup and view all the answers

Who, along with his team, first derived human embryonic stem cells in 1998?

James Thomson Signup and view all the answers

What is the historical significance of Alexander Maksimov's suggestion of the name 'stem cells'?

He suggested the name for cells that can differentiate into another cell type Signup and view all the answers

What are the general characteristics of stem cells?

1- self renewing for long time : keeping stemness stage 2- potency : Have the ability to differentiate into wider range of adult cells 3- Plasticity: Can differentiate into another cell type other than its original tissue 4- Relocation: can migrate from its original place to another place Signup and view all the answers

What are the different classifications of stem cells based on their potency?

Totipotent, Pluripotent, Multipotent, Oligopotent, Unipotent, Induced pluripotent Signup and view all the answers

Where do embryonic stem cells come from and what is their potency?

Embryonic stem cells come from embryos that are three to five days old and are pluripotent. Signup and view all the answers

Where are adult stem cells found and how do they compare with embryonic stem cells?

Adult stem cells are found in small numbers in most adult tissues, such as bone marrow or fat. Compared with embryonic stem cells, adult stem cells have a more limited ability to give rise to various cells of the body. Signup and view all the answers

How are induced pluripotent stem cells (ipSCs) created and what is their significance?

Induced pluripotent stem cells are created by transforming regular adult cells into stem cells using genetic reprogramming. Their significance lies in providing an alternative to using embryos and aborted babies for stem cell research. Signup and view all the answers

What are perinatal stem cells and where are they found?

Perinatal stem cells are found in amniotic fluid as well as umbilical cord blood. Signup and view all the answers

Name some examples of body fluids where stem cells are found.

Stem cells are found in synovial fluid, amniotic fluid, menstrual blood, and mother milk. Signup and view all the answers

In which cells can medical signaling cells help treat pain and protect body cells from death?

Medical signaling cells can help treat pain and protect body cells from death due to lack of Mesenchymal Stem Cells. Signup and view all the answers

What happens when a stem cell divides?

When a stem cell divides, each new cell has the potential to either remain a stem cell or become another type of cell with a more specialized function. Signup and view all the answers

How can stem cells be classified based on their sources?

Stem cells can be classified into embryonic or adult based on their sources. Signup and view all the answers

Study Notes

Applications of Cell Culture

Supports drug development through testing drug efficacy and toxicity on specific cell types.
Aids in vaccine production, as cultured cells can host viral antigens for vaccine formulations.
Facilitates research in cell biology, enabling studies of cellular processes and interactions.

Primary vs. Secondary Culture

Primary culture involves the initial isolation of cells from tissues, with cells directly obtained and maintained in culture conditions.
Secondary culture, or passage culture, comes from subculturing cells from a primary culture, expanding cell lines while maintaining specific characteristics.

Cryopreservation Steps

Prepare the cells by detaching them and resuspending in a freezing medium containing cryoprotectants like DMSO.
Freeze the cell suspension gradually using a controlled-rate freezer or insulated container to prevent ice crystal formation.
Store frozen cells in liquid nitrogen for long-term preservation.

Purpose and Steps of Subculture (Passaging)

Subculture is performed to maintain healthy cell growth and prevent over-confluence.
Steps include detaching cells from the culture vessel, re-suspending in fresh medium, and plating them into new culture vessels.

Challenges in DNA Purification

Endogenous nucleases can compromise DNA integrity by degrading nucleic acids.
Effective inactivation of nucleases can be achieved through heat treatment, using specific inhibitors, or maintaining low temperatures during extraction.

Definitions

Cell Culture: A process of growing cells in a controlled environment, typically outside their natural habitat.
Cell Strainer: A device used to separate cells based on size, allowing for the removal of clumps and debris.
Anchorage-Dependent Cells: Cells that require a surface to attach for growth and proliferation.
Anchorage-Independent Cells: Cells that can grow in suspension without needing a surface for attachment.

Basic Steps in DNA Extraction

Cell lysis to release DNA from the cell nucleus.
Separation of cellular debris from nucleic acids.
Precipitation and purification of DNA from the solution.

Role of Protease in DNA Extraction

Proteases are added to degrade proteins that could contaminate the nucleic acids, facilitating a purer DNA sample.

Treatment of Debris in DNA Extraction

Debris such as broken proteins, lipids, and RNA is separated through centrifugation or filtration, ensuring cleaner DNA isolation.

Purpose of Centrifugation in DNA Extraction

Centrifugation separates cellular debris from the lysate, allowing for clearer isolation of nucleic acids.

Ethanol Precipitation in DNA Extraction

Ethanol is used to precipitate DNA from the solution, promoting DNA recovery while removing impurities.

Minicolumn Purification Step

DNA isolation occurs as the sample is passed through a minicolumn, where DNA binds to the column matrix and contaminants are washed away.

Traditional Genomic DNA Isolation Method

The phenol-chloroform extraction method has been widely used for isolating genomic DNA from various sources.

Isoamyl Alcohol in Phenol Extraction

Isoamyl alcohol helps reduce foaming during the extraction process, improving the separation of aqueous and organic layers.

Removal of Proteins by Salting Out Method

High salt concentrations precipitate proteins, allowing for their separation from the DNA solution.

DNA Behavior During Salting Out

DNA remains soluble while proteins precipitate, enabling effective purification.

Origin of the Term 'Stem Cells'

Alexander Maksimov suggested the term "stem cells" in 1908 for cells capable of differentiating into various cell types.

Derivation of Embryonic Stem Cells

ES cells were first derived from mouse embryos in 1981 by researchers Martin Evans and Gail Martin.

Discovery of Mesenchymal Stem Cells

The discovery was made by Pittenger et al. in 1991, identifying a unique population of stem cells within the connective tissue.

First Human Embryonic Stem Cells

James Thomson and his team were the first to derive human embryonic stem cells in 1998.

Historical Significance of Stem Cells

Maksimov's naming highlights the potential of these cells in regenerative medicine and developmental biology.

General Characteristics of Stem Cells

Stem cells are characterized by their ability to self-renew, differentiate into specialized cell types, and respond to various stimuli.

Classifications of Stem Cells by Potency

Stem cells can be classified as totipotent, pluripotent, multipotent, and unipotent based on their differentiation potential.

Source and Potency of Embryonic Stem Cells

Embryonic stem cells are derived from the blastocyst stage of embryos and are pluripotent, able to differentiate into any cell type.

Location and Comparison of Adult Stem Cells

Adult stem cells are found in various tissues (e.g., bone marrow) and are typically multipotent, allowing them to give rise to a limited range of cells compared to embryonic stem cells.

Creation and Significance of Induced Pluripotent Stem Cells (iPSCs)

iPSCs are created by reprogramming adult somatic cells to an embryonic stem cell-like state, holding promise for personalized medicine and regenerative therapies.

Perinatal Stem Cells

Found in perinatal tissues, such as umbilical cord blood and placenta, these stem cells have unique properties for therapeutic use.

Sources of Stem Cells in Body Fluids

Stem cells can be found in body fluids including amniotic fluid, blood, and saliva.

Role of Medical Signaling Cells

Medical signaling cells assist in pain management and protect against cell apoptosis, enhancing tissue repair and regeneration.

Division of Stem Cells

When a stem cell divides, it can produce another stem cell (self-renewal) and a differentiated cell that will develop into a specific cell type.

Classification of Stem Cells by Source

Stem cells can be categorized based on their origin as either embryonic, adult, or induced stem cells.

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Description

Test your knowledge on cell culture applications and techniques with this quiz. Explore topics such as applications of cell culture, cryopreservation, primary and secondary culture, and cell subculture.