Xeno-free culture media-mediated rejuvenation of Hutchinson?Gilford Progeria Syndrome skin fibroblast

초록

Hutchinson?Gilford Progeria Syndrome (HGPS) is a rare genetic disorder caused by a mutation in the LMNA gene, which encodes lamin A, a protein crucial for maintaining the stability of the cell nucleus. This mutation produces an abnormal protein called progerin, which weakens the nuclear membrane and accelerates cellular aging, ultimately leading to premature aging of the body. Inhibiting progerin is a critical and validated strategy for delaying or stopping the pathological aging process in patients with HGPS. In this study, we cultured skin fibroblast cell lines from an HGPS patient using human platelet lysate-containing, xeno-free (XF) media. Interestingly, we observed a significant decrease in progerin expression compared to cells cultured in media containing fetal bovine serum (FBS). Consistently, we saw reduced cell size and increased proliferative capacity, indicated by a shorter population doubling time and a higher proportion of BrdU-positive cells. Additionally, p21 expression decreased at both the RNA and protein levels. RNA sequencing analysis revealed that genes strongly associated with the cell cycle, including EXO1, FANCA, FEN1, and UHRF1, were upregulated, whereas ID1 and GEM were downregulated. The percentage of cells displaying γH2AX foci was lower, while H3K9me3 and HP1γ levels increased in cells cultured with XF media, suggesting less DNA damage and improved genome stability and transcriptional regulation. Moreover, mitochondrial function improved, as evidenced by increased JC-1 staining and decreased production of mitochondrial reactive oxygen species. Finally, β-galactosidase activity declined, indicating reduced cellular senescence in XF media. In conclusion, XF media likely contains potential rejuvenating factors that may help slow the premature aging process in patients with Hutchinson-Gilford Progeria Syndrome. Further research is needed to identify and understand these factors.