masteroc Posted September 16, 2012 Posted September 16, 2012 I'm taking my first tissue culture course at university and I had a rather fundamental questions that I was hoping I could get some help with. The book that we are using does not like to define terms and seems (at least to me) rather difficult to follow. The book has sections on substrates, substrate preperation, buffered salt solutions, media selection, etc. What is really bothering me is that it doesn't really lay out the actual layers or components that you need to grow cells (lets say eukaryotic cell cultures to keep it simple). If this seems like an ambiguous question, what I mean is that it has said that substrates need to be treated to get cells to adhere (based on which substrate you choose) and then it says that cells will excrete their own ECM or at least components (such as collagen, fibronectin, gelatin). Then the book goes on to explain about buffered salt solutions and media selection, such as DMEM, MEM, etc. What really confuses me is just what is required to have these cell cultures grow. Lets say I wanted to start out with cell culture X, what are the layers that I need for these cells to take. So what I guess im asking for is this: what do you actually need for cell adhesion and proliferation in a cell culture. If you could keep it as introductory as possible (this is my first tissue culture course) that would be much appreciated. Thanks!
CharonY Posted September 19, 2012 Posted September 19, 2012 For the actual adhesion you just need a suitable surface (such as polystyrene, though in some cases you need an existing cell layer, for example). The cells will first settle on the surface, but to actually adhere and grow you need to keep the cells happy by providing a suitable environment (including temperature, pH, and gas atmosphere), as well as nutrients. Practically you have a medium in a suitable dish and seed it with fresh cells. The cells will settle, adhere, and form a layer within that dish (if you keep them happy).
mohamed samir Posted March 9, 2013 Posted March 9, 2013 First of all, Iam happy that you do your best to capture some valuable information from the book in your hand. Here and as far as I know that the surface of the culture bottle or the culture petri dishes characterized by presence of material, polystyrene. that help the cell to adhere to the surface and I think also when u provide the cell with nutrient and medium , after 1-2 hours it will start gowing and adhere to the surface
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