Dear all, can anyone tell me that
1. if the methylation level of some certain genes can change in cultured cells（without any special treatment ) or in stored DNA sample?
2. If the methylation level of some genes in a cell line must be 100% or 50% or 0%? If not, does it mean the cell line is not a real clone?
Here I mean single CpG site's methylation, not the average methylation level of a region.
Thanks in advance!
Some studies of methylation in human cell lines in culture have shown variations of methylation status, especially for cancer cell lines, for example for rRNA genes (hypomethylation in cancer cell lines with elevated transcription of these genes, hypermethylation in some cell lines where these genes are poorly expressed). I do not find report of methylation changes during cell culture
Methylation status of a gene(s) in cells depends on a number of factors, including the culture conditions. Therefore, it is not surprising that one finds differences. In part, it could be due to clonal variations in cell populations. Once DNA is isolated from cells and it is proteins-free, it is unlikely that the methylation status would change upon storage.
Methylation status changes in culture conditions.. it would be good if you want to really study it try to isolate DNA in differentnstages of cell passages and do Bisulphite sequencing ... costly affair ... we can do for you at Xcelris
Thank you all very much.
We're a gene test company and we are developing a methylation (of some gene) test kit recently. We need to confirm that the methylation status of DNA extracted from FFPE samples will not change during the storage.
Besides, since I was told that the DNA methylation level of cell line should be 0% or 50% or 100%, I wonder if it is normal that the methylation levels of the cell lines we used as reference materials are 60% or 80%.
Anyway, I'm inspired by your answers. Thank you again!
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