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Two Exam Cell Culturing Techniques
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What is needed for preparation of primary cell culture?
Enzymes
Protocols
Antibiotics
Primary culture for explants
What are the advantages and disadvantages of cell culture?
Advantage- Development of cells line over generations
Disadvantage- may lose differentiated characteristics
What are the most commonly used enzymes for disaggregating cells?
Trypsin and Pronase
What are the 2 types of continuous culture?
Cell lines
Continuous Cell lines
What is transformation?
Spontaneous or induced permanent phenotypic changes resulting from change in DNA and gene expression
What is transfection?
Introduction of DNA into a cells (like viral DNA)
What is the difference between histotypic and organotypic cultures?
Histo- single cell line
Organo- multiple cell lineages
What are the cell cycle phases and what happens in each?
G0/G1- pre-DNA synthesis
S- DNA synthesis
G2- period between DNA synthesis and mitosis
M- mitosis (actual cell division)
How do we measure cell growth?
Population doubling level (generation #)
Multiplication rate (r)
Population doubling time (PDT)
What is the equation for cell growth?
n= 3.32 (logNH- logN1)
N1- initial cells
NH- harvested cells
What is a cell line?
Cell population derived from a primary culture at the first passage
What is SOP (Standard Operating Procedure)?
Detailed instructions for technique used
All aspects of tasks involved
What are the 2 sub-zero temperatures used to cryopreserve cells?
-80
-196
Why freeze cells?
Reduce risk of microbial contamination, cross contamination, genetic drift and morphological changes
Reduce cost
What are the properties of Cryoprotectant?
Lowering freezing temperature
Forming Viscosity
Diffuse through membrane
What are conventional cryoprotectants?
glycols
What are the main factors the influence cell viability?
Cryoprotectant, cooling rate, storage, temperature, and warming rate
What are the advantages and disadvantages of a -135 freezer?
Pros- Easy to maintain
Steady Temp
Low running cost
Cons- Requires liquid nitrogen (LN) back up
Mechanically complex
Temperature is high compared to LN
What are the advantages and disadvantages of Liquid Phase Nitrogen?
Pros- Steady Ultra Low Temp
Simple and Mechanically reliable
Cons- Requires steady supply of LN
High Cost
Risk of cross-contamination
What are the pros and cons of Vapor Phase Nitrogen?
Pros- No risk of cross contamination
Low temp
Simplicity and reliability
Cons- Temperature fluctuation
Requires steady supply of liquid nitrogen
Costly
Author
icimmy
ID
66456
Card Set
Two Exam Cell Culturing Techniques
Description
Two Exam Cell Culturing Techniques (week four-six)
Updated
2011-02-15T13:47:52Z
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