Hello ! I am searching an in vitro model to study the implication of a protein in mitochondrial dysfunctions in dopaminergic cells.
Are SH-SY5Y the best model ? Are primary culture cells from mouse midbrain more suitable ?
SHSHY5Y IS A GOOD ONE !
THE PRIMARY NEURONS FROM MOUSE MIDBRAIN IS MORE SUITABLE, BUT I DONT THINK YOU CAN DO IT AS DA NEURONS IS HARD TO BE CULTURED.
Thank you for your point of view , Sun.
Do you have any idea on : - How to culture them. and - How many TH positive cells among all cells do you have ? 100% ?
You can use SH-SY5Y cells but there are better cell models than that. You can also use N27 cells and differentiated LUHMES cells for mitochondria dysfunction work
An important aspect to consider when assessing mitochondrial dysfunction in cultured cells is the metabolic state of the cells. It's important to use cells which actually utilize mitochondrial OXPHOS. Most cultured cells grown in glucose-containing medium are predominantly glycolytic and therefore not a good model for assessing mitochondrial dysfunction.
Here is a paper which describes a method for switching the metabolic state of cultured cells from glycolysis to OXPHOS. This switch increased susceptibility to the mitochondrial toxin rotenone.
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Thank you very much for your answers! N27 cells look very interesting, but I am hesitating because I read that :"Because the original N27 cell line has been passaged many times, the line has become a mixture of cell types with highly variable expression of TH" (Gao L et al, Plosone 2016). I am wondering if the commercially-available N27 cells have this problem...
Marie I am aware of that paper.. but that group have a commercial interest for that cells as the they sell the N27-A cells. In my personal experience if you get a N27 cells from good source or commercially, you would get pretty good TH expression as well as mitocondrial response you looking for. Also N27 cells very easy to maintain and to work with... something for you to think about..
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