Notesale: Turn your study into money

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Cell Culture - Basics
Primary Cell Culture- is the outgrowth of migrating cells from a piece of tissue, or tissue that has
been broken down by various methods such as Enzymatic methods (Trypsin) , Chemical methods
(EDTA) or mechanical methods (homogenisation)
Basically: primary cell culture are from cells freshly taken from a piece of tissue and broken down
Primary Cell Line – Cells that have attached after the desegregated process and proliferated
Cell cultures consist of purely eukaryotic cells (ie animal cells)

Adherent cells are cells that stick to a surface and multiply
...

It is worth noticing that the cells on the left grow in a
monolayer then stop dividing (this is called the hay flick
phenomenon) ; this rule applies for all adherent cells unless the
cell culture is cancerous
...
The main two examples of this are
Erythrocytes (red blood cells) and Leukocytes (white blood
cells)

A cell suspension media consists of:

Foetal Calf Serum:
This is the plasma from
centrifuged calf’s blood

Penicillin:
To remove any Gram positive
bacteria that may have
contaminated the media

5% CO2:
To induce proliferation

Remember how messy you got when you were a kid and got paint on your hand – well the same
principle applies in cell cultures, but instead of paint there’s microbial contamination!

Cell cultures must be performed in sterile conditions as there is no immunity to protect the dividing
cells from the microbes; this is why the correct equipment is essential!
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Mask – to stop you exhaling on the sample and contaminating it (there’s bacteria in your
mouth too you know!)
Gloves – To protect from microbial contamination transferred from your hands
Overalls – to protect from microbial contamination from your clothing
Air hood – to filter and circulate air to remove any contamination
70% Ethanol – to sterilise any equipment and environment

Finally:
You may be wondering why I’m using the word microbial rather than bacterial contamination, this is
because few people realise more than bacteria can contaminate the sample

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