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Cell Culture

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musselburghgrammar

The document discusses mammalian cell culture and its applications. It provides details on the conditions needed for cell culture, including aseptic conditions, growth medium, temperature, pH, and gas exchange. It also discusses primary and continuous mammalian cell cultures. Primary cultures have a limited lifespan while continuous cell lines can divide indefinitely and are used for research and biotechnology applications.

Technology
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Mammalian Cell Culture ,[object Object]
Applications of cell culture 1 ,[object Object],[object Object],[object Object],[object Object]
Applications of cell culture 2 ,[object Object],[object Object],[object Object],[object Object],[object Object]
Conditions needed for cell culture ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Aseptic conditions ,[object Object],[object Object],[object Object],e.g. sterilisation of all utensils and media using heat . For example, using an autoclave (steam under pressure, necessary for bacterial spores) Growth of pure cultures
Microorganisms ,[object Object],[object Object],[object Object],[object Object]
Classes of Microorganisms ,[object Object],1. Unicellular Algae / PHOTOTROPHS : use sunlight to make their own food 1. Bacteria & Yeasts (Fungi) / HETEROTROPHS : need more complex media containing an organic carbon source and other compounds e.g. amino acids
Culture & Uses ,[object Object],[object Object],[object Object],[object Object],[object Object],[See Scholar: Batch & Continuous culture]
Microbial Growth Culture Requirements ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Microbial Growth Culture Requirements ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Unicellular algae , bacteria & yeast can be grown as batch cultures - no dilution is needed until max. density is reached. Growth can be limited by nutrient availability i.e. at the end of exponential growth
Nutrient Media ,[object Object],[object Object],[object Object],[object Object]
Nutrient Media ,[object Object],1. Complex media - this has one or more crude sources of nutrients and their exact chemical composition and components are not known. Generally used for routine cultures 2. Defined media - otherwise known as synthetic media containing chemically known compounds and components which are in a relatively pure form REMEMBER: all media must be STERILE before use !!!
Mammalian Cell Culture ,[object Object]
The process of culturing Mammalian Cells ,[object Object],The cells stick or adhere to the inside of the vessel as they grow in the medium Most animal cells are ‘ANCHORAGE-DEPENDENT’ i.e. they need something to hold on to These cells usually form a monolayer that will eventually cover the entire surface of the medium
At this point, called confluence , it is necessary to subculture the cells into a fresh medium N.B. Cells that are associated with body fluids such as blood cells are NON-ANCHORAGE DEPENDENT and can be grown in suspension. Again, it is necessary to regularly subculture the cells into fresh medium N.B. All media and culture vessels are STERILISED to prevent the growth of micro-organisms
Mammalian Cell Growth Media ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Categories Of Mammalian Cell Cultures ,[object Object],[object Object],[object Object],[object Object]
Process of Cell Collection ,[object Object],[object Object],[object Object]
[object Object],[object Object],[object Object],[object Object]
Continuous Cell Lines ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Animal Cell Culture Solutions fragile cells ; can’t be stirred air lift fermenters slow growth ; risk of contamination Problems aseptic transfer in sterile air flow need complex nutrients media with calf serum etc. ; can’t be heat sterilised media sterilised by filtration high contamination risk
Bacterial & Fungal Cultures ,[object Object],[object Object],[object Object]

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Cell Culture

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  • 15. At this point, called confluence , it is necessary to subculture the cells into a fresh medium N.B. Cells that are associated with body fluids such as blood cells are NON-ANCHORAGE DEPENDENT and can be grown in suspension. Again, it is necessary to regularly subculture the cells into fresh medium N.B. All media and culture vessels are STERILISED to prevent the growth of micro-organisms
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  • 21. Animal Cell Culture Solutions fragile cells ; can’t be stirred air lift fermenters slow growth ; risk of contamination Problems aseptic transfer in sterile air flow need complex nutrients media with calf serum etc. ; can’t be heat sterilised media sterilised by filtration high contamination risk
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