Cell Culture
Types of Cell culture
A starter culture of microbial cells is inoculated & grown in/on:
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1. Broth (liquid media) ​​​​ 2. Agar (solid media)
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Nutrients in Cell Culture
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Culture media contains suitable nutrients that promote the growth of specific cells/ microbes.
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Animal cells
Grown in media containing growth factors from serum.
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Growth factors= Proteins that promote cell growth/proliferation.
Aseptic Technique
Involves the sterilisation of equipment/culture media when carrying out cell culture experiments by heat or chemicals.
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Importance
Eliminates unwanted microbial contaminants when culturing microbes.
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heat chemicals
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Measuring Cell Growth
1. Colony forming units (CFU)
Plating out of a liquid microbial culture on solid media allows the number of CFU to be counted and the density of cells in the culture estimated.
Serial dilution is often needed to achieve a suitable colony count
(30 to 300 colonies).
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Measuring Cell Growth
2. Haemocytometer with Vital Staining
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​Use of haemocytometer to estimate cell numbers per cm3 in a liquid
broth culture.
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​​​​​​​​​​​​​​​​​​​Vital Staining of Cells
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Measuring viable cell count on haemocytometer
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Use of stains such as trypan/methylene blue that only stain dead cells as viable cells remove blue dye by active transport.
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Measuring Cell Growth
