I am struggling to get fine cholesterol crystal for cell.
I have got cholesterol crystal through the methods from many papers, but I cannot get very fine particles from relatively bigger particles.
Simply, I choose to utilize sterile Mortar and pestle to grind. It does not seem to be small particle enough to apply to cell. FYI, I am using macrophage cell lines. Also, cells are primed with LPS ,then stimulated with Cholesterol crystal.
I am looking forward to get a advise from you, expertise!
If you have access to a small volume ball mill, you could sterilise the parts and mill the cholesterol this way. This way you can get a very fine powder (I used this method to get homogenous samples of freeze-dried silage which is very fibrous still and it worked wonders).
Good luck!
Dear Yongeum,
You will not get cholesterol crystals into any aqueous solution i.e. cell culture media without changing its lipophilic character. You can either compelx it by your self with Methyl-β-cyclodextrin (publications explaining that are out there but old) but the qualitiy might vary or you can (and that is what I would advise) buy Cholesterol-Water Soluble from Sigma (C4951). That worked for me and it is possible to monitor it integration by lipid droplet staining.
Best Soenke
@Sabine Harrison
Thank you for your reply! It will be helpful for me. You think the small ball mill is the only way to get very fine powder?
@Sönke Weinert
I appreciate your answer. I already got cholesterol crystal through procedure using cholesterol power from Sigma. Since I want to study that cholesterol crystal and inflammation on the macrophage, I try to get crystal power. Do you think cholesterol water soluble cholesterol affect on inflammation procedure will be same with crystal?
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