Cell Biology and Genetics Lab Session Plan

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Questions and Answers

What is the primary focus of the laboratory session described?

Synthesis of artificial hormones

Maintenance and propagation of human immortalized cell lines (correct)

Analysis of plant cell structures

Development of organic compounds

What does the process of dissociating cells from compact tissue typically involve?

Application of heat to promote cell division

Freezing cells to separate them

Exposure to high-frequency sound waves

Use of proteolytic enzymes to disrupt adhesive bonds (correct)

What tool is mentioned for isolating specific types of cells from a tissue sample?

Electrophoresis chamber

Cryostat

Laser scanning microscope

Fluorescence-activated cell sorter (correct)

What is one of the applications of cell-based applications mentioned in the session?

Biochemical studies and vaccine development Signup and view all the answers

During the lab session, what general technique will students NOT be practicing?

Harvesting energy from cell cultures Signup and view all the answers

How long is the duration of the laboratory session for each group?

2 hours Signup and view all the answers

What is a key technique students will learn in this laboratory session related to handling cancer cells?

Splitting and re-plating of cancer cells Signup and view all the answers

What is the role of adhesive bonds between cells in tissue that may need to be disrupted?

To facilitate cell separation during dissociation Signup and view all the answers

Which factor is NOT an element of aseptic technique?

Use of contaminated reagents Signup and view all the answers

At what temperatures should cells NOT be stored to maintain viability?

-20°C and -80°C Signup and view all the answers

What is an advantage of using vapor-phase liquid nitrogen storage systems?

Minimized risk of explosion Signup and view all the answers

Which factor is NOT considered critical when purchasing a biosafety cabinet for cell culture?

Cost of the unit Signup and view all the answers

Why should sterile pipettes be handled with care?

To prevent cross-contamination Signup and view all the answers

When selecting a cell culture incubator, which parameter is least important for cell growth?

Oxygen levels Signup and view all the answers

Which type of storage container typically has a slower nitrogen evaporation rate?

Narrow-necked containers Signup and view all the answers

Which of the following is NOT a critical feature of a centrifuge used in cell culture?

Contamination prevention measures Signup and view all the answers

What is a key reason for preparing working stocks of cell lines?

To maintain genetic stability Signup and view all the answers

What is the main consideration when choosing between vapor-phase and liquid-phase storage systems?

Risk of explosion and static holding times Signup and view all the answers

What is considered the main benefit of using Fetal Bovine Serum (FBS) in cell culture media?

It enhances cell health and viability. Signup and view all the answers

Which equipment is primarily used to visualize cells in a culture?

Inverted microscope Signup and view all the answers

Why should the outside of containers be wiped with 70% ethanol?

To reduce the probability of contamination Signup and view all the answers

Which item is NOT typically considered an additional supply needed for cell culture?

Spectrophotometer Signup and view all the answers

What is the main reason for maintaining aseptic techniques in cell culture?

To reduce contamination from external microorganisms Signup and view all the answers

Which parameter is NOT directly influenced by the composition of cell culture media?

Contamination of samples Signup and view all the answers

What feature ensures that somatic cells do not divide indiscriminately and develop into cancerous cells?

Finite number of cell divisions Signup and view all the answers

What is the role of telomerase in somatic cells?

To maintain the ends of chromosomes during cell division Signup and view all the answers

What distinguishes immortalized cell lines from normal somatic cells?

Ability to divide indefinitely Signup and view all the answers

Which type of cell lines can give rise to any tissue in the body when appropriately treated?

Human embryonic stem (ES) cell lines Signup and view all the answers

What is a common requirement for all cell culture laboratories?

Asepsis or being free from pathogenic microorganisms Signup and view all the answers

In vitro experiments are performed under which conditions compared to in vivo experiments?

In controlled laboratory settings using cultured cells Signup and view all the answers

For a mammalian cell culture laboratory specializing in cancer research, which equipment would likely be essential?

Contamination detection systems Signup and view all the answers

What is the primary reason for the progressive shrinkage of chromosomes in human somatic cells?

Absence of telomerase expression Signup and view all the answers

What should be done with the cap or cover of a sterile flask before placing it down?

Place it with the opening facing down. Signup and view all the answers

Which of the following actions is crucial for maintaining sterility during experiments?

Keeping all containers covered until needed. Signup and view all the answers

What is the most appropriate method for adding wash solution to a culture vessel?

Gently add it to the side opposite the cell layer. Signup and view all the answers

What should be done immediately after removing the used cell culture media?

Ensure the caps or covers face down on the work area. Signup and view all the answers

What is the incubation time range for the dissociation reagent trypsin?

10-15 minutes Signup and view all the answers

Which procedure should be performed using the pipette during the washing of cells?

Ensure the pipette tip does not touch anything non-sterile. Signup and view all the answers

What should be done with the work surface before leaving the lab?

Wipe it with 70% ethanol. Signup and view all the answers

What is the appropriate action when transferring cells into a 6-well plate?

Ensure you label the plate with your group number. Signup and view all the answers

Study Notes

Laboratory Session Plan: Principles of Cell Biology and Genetics

Course: DES115 Dentistry
Module: Introduction to cell culture: cell plating, splitting, and freezing
Week: 8-9
Year: 1
Semester: 1
Lab Location: N41, N42
Lab Session Duration: 2 hours x 8 groups
Lab Instructor: Dr Harous C. and Dr Tomouzou C.
Learning Objectives: Teach students how to maintain and propagate human immortalized cell lines. To stress cell-based applications in basic research, biochemical studies, vaccine development, and drug discovery.
Lab Activities: Culturing human cancer cells, splitting, re-plating, and freezing
Instructor Instructions: Students will be assigned to teaching stations and link the techniques used in the lab to theoretical concepts taught in lectures. Cell culture, splitting, re-plating, and freezing human cancer cells will be performed during the session.

Introduction to Cell Culture

Cell Isolation: Various methods can separate specific cell types from tissues, including enzyme treatment to break down connections between cells, and cell sorting via equipment like a fluorescence-activated cell sorter (FACS).
In vitro vs. In vivo: Cell cultures are conducted in vitro (in glass/culture) to distinguish them from experiments on whole organisms (in vivo).
Immortalization: Some cells can divide indefinitely. This occurs due to a genetic change and is accomplished by adding the telomerase gene. These immortalized cell lines are a useful source of homogenous cells for study
Human Embryonic Stem (ES) Cells: An important type of cell line; ES cells are undifferentiated, meaning they can become any tissue type in the body. These hold potential for disease treatments

Cell Morphology

Fibroblastic/Fibroblast-Like Cells: Elongated shapes, bipolar or multipolar, attached to substrates, and grow attached to a surface.
Epithelial-Like Cells: Irregular or polygonal shape, grow attached to substrates in patches.
Lymphoblast-Like Cells: Spherical, and generally grow in suspension, not attached to substrates

Cell Culture Equipment

Cell Culture Hoods (Laminar Flow Hoods or Biosafety Cabinets): Control airflow for sterile work; essential for protecting samples and workers
Incubators (Humid CO2 Incubators): Maintain temperature, humidity, and CO2 levels for optimal cell growth.
Water Baths: Used for temperature control.
Centrifuges: Separate or concentrate cells; maintain precise speeds and temperature control.
Refrigerators and Freezers: Storage of equipment and supplies.
Cell Counters: Count cell numbers.
Inverted Microscopes: Observe cells in culture.
Liquid Nitrogen Freezers/Cryostorage Containers: Long-term storage of cells.
Sterilizers (Autoclaves): Sterilize equipment.

Additional Supplies

Cell culture media: Provides nutrients and growth factors necessary for cell growth and function.
Fetal Bovine Serum (FBS): Widely used serum to maintain and preserve cell health and viability.
Aseptic Technique: Crucial for preventing contamination of cell cultures by microorganisms.
Cryogenic Storage: Methods for long-term preservation of cells.

Experimental Procedure

Cell Handling: Proper disinfection practices and sterile technique are essential
Media and Reagent Handling: Avoid pouring directly from bottles; use sterile pipettes and instruments.
Cell Dissociation: Cells grown in culture may be detached employing trypsin solutions or other enzymes.
Cell Cultivation: Proper methods for incubating cells will be applied

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Description

This lab session plan for DES115 Dentistry covers the principles of cell culture, including techniques of cell plating, splitting, and freezing. Students will learn to maintain and propagate human immortalized cell lines, focusing on applications in research, vaccine development, and drug discovery.