Hello! I am pretty new to this kind of topic (lisosomes and artificial membranes in general) and I was wandering if anyone here knows or can suggest me some protocol in order to get this kind of structures starting from living cells. The idea would be to have something as similar as possible to the cell membrane.
Many thanks
Giovanni.
Hi Giovanni,
I guess you want to make liposomes. You can do that by mixing different mol% of various lipids in organic solvent followed by removing the solvent and rehydrating them in buffer of choice. After that you need to make the size of liposomes homogeneous by extrusion. Then you are ready to go!! I hope I answered your question.
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Best,
Kalpana
Hi Kalpana. Thanks for your answer but maybe I was not clear in my previous message.
I really wanted to do liposomes out of membranes deriving from actual cells. Do you know any protocol that would allow me to do something like that?
Hi Giovanni,
For that you will need to fractionate the cell membrane using lysis and ultracentrifugation or may be you have to fractionate membrane lipids. That will be your starting point. I will try to get you more information on this.
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Best,
Kalpana
Do you want them to contain the actual proteins as well? I think you can find articles where people have prepared giant unilamellar vesicles (GUVs) from RBC ghosts, and in principle cell membranes of any cell could be used, although it is quite a lot of work to set up such systems. See this paper by Luis Bagatolli, e.g.: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2072068/
If you are happy with small vesicles, you can use sonication to break down (the preferably isolated) cell membranes that typically with sufficient power produces small unilamellar vesicles (
Many thanks! I think the article you sent me is exactly what I was looking for.
Many thanks again, you saved my life!
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